U.S. patent application number 09/907581 was filed with the patent office on 2002-01-03 for devices, methods and systems for collecting material from a breast duct.
Invention is credited to He, Xuanmin, Hung, David, Ken, Christopher G.M., Nikolchev, Julian, O'Leary, Shawn, Olsen, Phillip M., Sayavong, Pam.
Application Number | 20020002343 09/907581 |
Document ID | / |
Family ID | 27537371 |
Filed Date | 2002-01-03 |
United States Patent
Application |
20020002343 |
Kind Code |
A1 |
Hung, David ; et
al. |
January 3, 2002 |
Devices, methods and systems for collecting material from a breast
duct
Abstract
The invention provides methods, devices and systems for
collecting breast ductal fluid comprising cellular material and
other useful markers for analysis. The methods typically comprise
access of at least one breast duct and collecting materials from
that duct separate from all other ducts in the breast. The devices
comprise ductal access devices that provide the opportunity to
collect fluid from a single duct separate from all the other ducts
in the breast. The systems employ the methods and devices that used
together provide systems for analysis of a breast condition in a
patient specific to accessed breast ducts. The methods, devices and
systems are particularly useful for indentification of breast
precancer or cancer in patient.
Inventors: |
Hung, David; (Belmont,
CA) ; Ken, Christopher G.M.; (San Mateo, CA) ;
He, Xuanmin; (Palo Alto, CA) ; Olsen, Phillip M.;
(Mountain View, CA) ; Nikolchev, Julian; (Portola
Valley, CA) ; O'Leary, Shawn; (San Jose, CA) ;
Sayavong, Pam; (Newark, CA) |
Correspondence
Address: |
BANNER & WITCOFF
1001 G STREET N W
SUITE 1100
WASHINGTON
DC
20001
US
|
Family ID: |
27537371 |
Appl. No.: |
09/907581 |
Filed: |
July 19, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09907581 |
Jul 19, 2001 |
|
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09473510 |
Dec 28, 1999 |
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60114048 |
Dec 28, 1998 |
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60134613 |
May 18, 1999 |
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60143476 |
Jul 12, 1999 |
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60143359 |
Jul 12, 1999 |
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60170997 |
Dec 14, 1999 |
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Current U.S.
Class: |
600/573 |
Current CPC
Class: |
A61M 3/0283 20130101;
A61M 3/0287 20130101; A61M 3/0262 20130101; A61M 2210/1007
20130101; A61B 10/0045 20130101; A61B 10/0041 20130101; A61M 1/88
20210501; A61M 3/022 20140204; A61M 3/0279 20130101; A61M 1/0062
20130101; A61M 1/815 20210501; G01N 33/57415 20130101 |
Class at
Publication: |
600/573 |
International
Class: |
A61B 005/00 |
Claims
What is claimed is:
1. A method for obtaining cellular material from a human breast
milk duct, said method comprising: introducing a wash fluid to the
breast milk duct, wherein a volume of at least 2 ml is present
within the duct for a preselected time; and collecting at least a
portion of the introduced wash fluid from within the duct, wherein
said portion carries the cellular material.
2. A method as in claim 1, wherein the preselected time is less
than one second.
3. A method as in claim 1, wherein the preselected time is in the
range from one second to one hour.
4. A method as in claim 1, wherein the wash fluid is introduced to
a volume of at least 2 ml prior to collecting any of wash fluid
from the duct.
5. A method as in claim 1, wherein the wash fluid is introduced to
a single breast milk duct and collected from the same breast milk
duct without mixing with materials from other breast milk
ducts.
6. A method as in claim 1, further comprising massaging and
squeezing the breast tissue after introducing the wash fluid but
prior to and during collecting a portion of the wash fluid.
7. A method as in claim 1, further comprising separating cellular
material from the collected fluid.
8. A method as in claim 7, further comprising examining the
separated cellular material.
9. A method as in claim 1, wherein the cellular material is a
substance selected from the group consisting of whole cells,
cellular debris, proteins, nucleic acids, polypeptides,
glycoproteins, lipids, fats, glycoproteins, small organic
molecules, metabolites, and macromolecules.
10. A method as in claim 1, wherein the wash fluid is selected from
the group consisting of saline, phosphate buffered saline, a
nonabsorbable fluid, an isotonic solution, an osmotic solution, a
hypotonic solution, and a hypertonic solution.
11. A method as in claim 1, wherein the wash fluid is selected from
the group consisting of a protein, a colloid, a sugar, a polymer,
mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, polyethyleneglycol (PEG),
maltodextrin, dextran (e.g. dextran 70), hydroxyethyl starch, fluid
gelatin, a synthetic colloid, an antibody, a binding protein, and
albumin.
12. A method for obtaining cellular material from a human breast
milk duct, said method comprising: introducing a ductal access
device having at least one lumen therethrough into a duct;
introducing a wash fluid through the access device lumen into the
milk duct, wherein a volume of at least 2 ml is present within the
duct for a preselected time; and collecting at least a portion of
the wash fluid from the duct through the lumen of the access
device.
13. A method as in claim 12, further comprising massaging and
squeezing the breast tissue after introducing the wash fluid but
prior to and during collecting a portion of the wash fluid.
14. A method as in claim 12, wherein introducing the ductal access
device comprises positioning a distal end thereof distal to the
ductal sphincter.
15. A method as in claim 12, wherein the access device has only a
single lumen which extends into the duct.
16. A method as in claim 12, wherein the wash fluid is introduced
to a volume of at least 2 ml prior to collecting any of wash fluid
from the duct.
17. A methods as in claim 12, wherein the preselected time is less
than one second.
18. A method as in claim 12, wherein the preselected time is in the
range from one second to one hour.
19. A method as in claim 12, wherein the wash fluid is introduced
to a single breast milk duct and collected from the same breast
milk duct without mixing with materials from other breast milk
ducts.
20. A method as in claim 12, further comprising separating cellular
material from the collected fluid.
21. A method as in claim 20, further comprising examining the
separated cellular material.
22. A method as in claim 17, wherein the cellular material is a
substance selected from the group consisting of whole cells,
cellular debris, proteins, nucleic acids, polypeptides,
glycoproteins, lipids, fats, glycoproteins, small organic
molecules, metabolites, and macromolecules.
23. A method as in claim 12, wherein the wash fluid is selected
from the group consisting of saline, phosphate buffered saline, a
nonabsorbable fluid, an isotonic solution, an osmotic solution, a
hypotonic solution, and a hypertonic solution.
24. A method as in claim 12, wherein the wash fluid is selected
from the group consisting of a protein, a colloid, a sugar, a
polymer, mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, polyethyleneglycol (PEG),
maltodextrin, dextran (e.g. dextran 70), hydroxyethyl starch, fluid
gelatin, a synthetic colloid, an antibody, a binding protein, and
albuinin.
25. A method for obtaining cellular material from a human breast
milk duct, said method comprising: introducing a wash fluid to the
breast milk duct, wherein the wash fluid is present within the duct
for a preselected time; and collecting at least a portion of the
introduced wash fluid from within the duct, wherein said portion
carries the cellular material; wherein the wash fluid is introduced
to a single breast milk duct and collected from the same breast
milk duct without mixing with materials from other breast milk
ducts.
26. A method as in claim 25, wherein the volume of wash fluid is at
least 2 ml.
27. A method as in claim 25, wherein the wash fluid is introduced
to a volume of at least 2 ml prior to collecting any of wash fluid
from the duct.
28. A method as in claim 25, wherein the wash fluid is introduced
to a volume of at least 2 ml prior to collecting any of wash fluid
from the duct.
29. A method as in claim 25, further comprising massaging and
squeezing the breast tissue after introducing the wash fluid but
prior to and during collecting a portion of the wash fluid.
30. A method as in claim 25, wherein the wash fluid is selected
from the group consisting of saline, phosphate buffered saline, a
nonabsorbable fluid, an isotonic solution, an osmotic solution, a
hypotonic solution, and a hypertonic solution.
31. A method as in claim 25, wherein the wash fluid is selected
from the group consisting of a protein, a colloid, a sugar, a
polymer, mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, polyethyleneglycol (PEG),
maltodextrin, dextran (e.g. dextran 70), hydroxyethyl starch, fluid
gelatin, a synthetic colloid, an antibody, a binding protein, and
albumin.
32. A method as in claim 25, wherein the preselected time is less
than one second.
33. A method as in claim 25, wherein the preselected time is in a
range from one second to one hour.
34. A method as in claim 25, further comprising separating cellular
material from the collected fluid.
35. A method as in claim 34, further comprising examining the
separated cellular material.
36. A method as in claim 25, wherein the cellular material is a
substance selected from the group consisiting of whole cells,
cellular debris, nucleic acids, lipids, protein metabolites, small
organic molecules, and macromolecules.
37. A method for obtaining cellular material from a human breast
milk duct, said method comprising: introducing a ductal access
device having at least one lumen therethrough into a duct;
introducing a wash fluid through the access device lumen into the
milk duct, wherein the wash fluid is present within the duct for a
preselected time; and collecting at least a portion of the wash
fluid from the duct through the lumen of the access device; wherein
the wash fluid is introduced to a single breast milk duct and
collected from the same breast milk duct without mixing with
materials from other breast milk ducts.
38. A method as in claim 37, wherein the volume of wash fluid is at
least 2 ml.
39. A method as in claim 37, wherein the wash fluid is introduced
to a volume of at least 2 ml prior to collecting any of wash fluid
from the duct.
40. A method as in claim 37, wherein the wash fluid is introduced
to a volume of at least 2 ml prior to collecting any of wash fluid
from the duct.
41. A method as in claim 37, wherein the preselected time is less
than one second.
42. A method as in claim 37, wherein the preselected time is in a
range from one second to one hour.
43. A method as in claim 37, further comprising massaging and
squeezing the breast tissue after introducing the wash fluid but
prior to and during collecting a portion of the wash fluid.
44. A method as in claim 37, wherein introducing the ductal access
device comprises positioning a distal end thereof distal to the
ductal sphincter.
45. A method as in claim 37 wherein the access device has only a
single lumen which extends into the duct.
46. A method as in claim 37, further comprising separating cellular
material from the collected fluid.
47. A method as in claim 46, further comprising examining the
separated cellular material.
48. A method as in claim 37, wherein the cellular material is a
substance selected from the group consisiting of whole cells,
cellular debris, nucleic acids, lipids, protein metabolites, small
organic molecules, and macromolecules.
49. A method as in claim 37, wherein the wash fluid is selected
from the group consisting of saline, phosphate buffered saline, a
nonabsorbable fluid, an isotonic solution, an osmotic solution, a
hypotonic solution, and a hypertonic solution.
50. A method as in claim wherein the wash fluid is selected from
the group consisting of a protein, a colloid, a sugar, a polymer,
mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, polyethyleneglycol (PEG),
maltodextrin, dextran (e.g. dextran 70), hydroxyethyl starch, fluid
gelatin, a synthetic colloid, an antibody, a binding protein, and
albumin.
51. A kit comprising: a ductal access device; and instructions for
use setting forth a method according to claim 12.
52. A ductal access device comprising: an access tube having a
distal end, at least one lumen therethrough, and dimensions which
permit introduction of the distal end through a ductal orifice and
positioning a distal end thereof distal to the ductal sphincter of
a human breast.
53. A ductal access device as in claim 52, further comprising means
on the access tube for positioning the distal end distal to the
ductal sphincter.
54. A ductal access device as in claim 53, wherein the positioning
means comprises length indicia on the tube which permit a user to
determine the depth to which the distal end of the tube has been
introduced.
55. A ductal access device as in claim 53, wherein the positioning
means comprises a stop element formed or attached to the tube,
wherein the stop has dimensions which prevent further insertion of
the tube into the duct and wherein the stop is positioned on the
tube so that the distal tip will be located distal to the ductal
sphincter when the device is fully inserted up to the stop.
56. A ductal access device as in claim 55, wherein the stop element
comprises a collar affixed to or formed on an exterior surface of
the tube.
57. A ductal access device as in claim 52, further comprising means
for anchoring the device to the breast.
58. A ductal access device as in claim 52, further comprising a
receiving portion comprising a water tight seal for receiving the
dilator.
59. A ductal access device as in claim 55, wherein the stop element
comprises a hub attached to a proximal end of the tube, wherein the
hub has a width which is greater than the diameter of the tube so
that a shoulder is formed at a junction between the tube and the
hub.
60. A ductal access device as in claim 53, wherein the positioning
means comprises a nob on the access tube having an increase
diameter for anchoring the tube distal to the ductal sphincter once
the nob has passed the sphincter and rests distal to it.
61. A ductal access device as in claim 52, wherein the access tube
has an outer diameter of 0.05 inches or 1.27 mm or less.
62. A ductal access device as in claim 52, wherein the access tube
has an outer diameter of 0.010 inches (or 0.254 mm) or greater.
63. A ductal access device as in claim 53, wherein the outer
diameter is in the range from 0.010 inches or 0.254 mm to 0.050
inches or 1.27 mm.
64. A ductal access device as in claim 52, wherein the access tube
has a lumen diameter 0.007 inches (or 0.178 mm) or greater.
65. A ductal access device as in claim 52, wherein the access tube
has a lumen diameter in the range from 0.007inches or 0.178 mm to
0.047 inches or 1.19 mm.
66. A ductal access device as in claim 52, further comprising: an
infusion connector providing a fluid flow path into the lumen of
the tube; and a collection connector providing a fluid outlet path
from the lumen of the tube, said infusion and collection connectors
being isolated from each other so that the fluid may be infused
through the infusion connector and simultaneously removed through
the collection connector.
67. A ductal access device as in claim 52, further comprising a
dilator removably received in the access tube and having a distal
tip which is positionable through the access tube to extend from
the distal end thereof.
68. A ductal access device as in claim 67, wherein the dilator has
an outer diameter of 0.024 inches (or 0.61 mm) or less.
69. A ductal access device as in claim 67, wherein the dilator is
tapered.
70. A ductal access device as in claim 67, wherein a receiving
portion of the device for receiving the dilator comprises a
water-tight seal.
71. A ductal access system comprising: a ductal access device as in
claim 52; and a container holding a premeasured volume of ductal
wash fluid.
72. A ductal access system as in claim 71, wherein the container
comprises a syringe for connection to the first side port.
73. A ducal access system as in claim 71, wherein the pre-measured
volume is in the range from 2 ml to 100 ml.
74. A ductal access system as in claim 71, wherein the ductal
access fluid is selected from the group consisting of saline,
phosphate buffered saline, a nonabsorbable fluid, an isotonic
solution, an osmotic solution, a hypotonic solution, and a
hypertonic solution.
75. A ductal access system as in claim 71, wherein the ductal
access fluid is selected from the group consisting of a protein, a
colloid, a sugar, a polymer, mannitol, sorbitol, glucose, glycerol,
sucrose, raffinose, fructose, lactulose, sodium chloride,
polyethyleneglycol (PEG), maltodextrin, dextran (e.g. dextran 70),
hydroxyethyl starch, fluid gelatin, a synthetic colloid, an
antibody, a binding protein, and albumin.
76. A ductal access device comprising: an access tube having a
distal end, a single lumen therethrough, and dimensions which
permit introduction of the distal end through a ductal orifice and
positioning a distal end thereof distal to the ductal sphincter. an
infusion connector providing a fluid flow path into the lumen of
the access tube; and a collection connector providing a fluid
outlet path from the lumen of the access tube, said infusion and
collection connectors being isolated from each other so that fluid
may be infused through the infusion connector and simultaneously
removed through the collection connector.
77. A ductal access device as in claim 76, wherein the tube has an
outer diameter of 0.010 inches or 0.254 mm or greater.
78. A ductal access device as in claim 76, wherein the tube has an
outer diameter of 0.050 inches or 1.27 mm or less.
79. A ductal access device as in claim 77, wherein the outer
diameter is in the range from 0.010 inches or 0.254 mm to 0.050
inches or 1.27 mm.
80. A ductal access device as in claim 76, wherein the access tube
has a lumen diameter 0.007 inches or 0.178 mm or greater.
81. A ductal access device as in claim 79, wherein the access tube
has a lumen diameter in the range from 0.007 inches or 0.178 mm to
0.047 inches or 1.1.9 mm.
82. A ductal access device as in claim 76, further comprising means
on the access tube positioning a distal end thereof distal to the
ductal sphincter.
83. A ductal access device as in claim 82, wherein the positioning
means comprises length indicia on the tube which permit a user to
determine the depth to which the distal end of the tube has been
introduced.
84. A ductal access device as in claim 82, wherein the positioning
means comprises a stop element formed or attached to the tube,
wherein the stop has dimensions which prevent further insertion of
the tube into the duct and wherein the stop is positioned on the
tube so that a distal end of the distal tip is positioned thereof
distal to the ductal sphincter.
85. A ductal access device as in claim 84, wherein the stop element
comprises a collar affixed to or formed on an exterior surface of
the tube.
86. A ductal access device as in claim 84, wherein the stop element
comprises a hub attached to a proximal end of the tube, wherein the
hub has a width which is greater than the diameter of the tube so
that a shoulder is formed at a junction between the tube and the
hub.
87. A ductal access device as in claim 82, wherein the positioning
means comprises a nob on the access tube having an increase
diameter for anchoring the tube distal to the ductal sphincter once
the nob has passed the sphincter and rests distal to it.
88. A ductal access device as in claim 76, further comprising means
for anchoring the device to the breast.
89. A ductal access device as in claim 76, further comprising a
dilator removably received in the access tube and having a distal
tip which is positionable through the access tube to extend from
the distal end thereof.
90. A ductal access device as in claim 89, wherein the dilator has
an outer diameter of 0.024 inches (or 0.061 mm) or less.
91. A ductal access device as in claim 89, wherein the dilator is
tapered.
92. A ductal access device as in claim 89, wherein a receiving
portion of the device for receiving the dilator comprises a
water-tight seal.
93. A ductal access system comprising: a ductal access device as in
claim 76; and a container holding a premeasured volume of ductal
wash fluid.
94. A ductal access system as in claim 93, wherein the ductal wash
fluid is selected from the group consisting of a protein, a
colloid, a sugar, a polymer, mannitol, sorbitol, glucose, glycerol,
sucrose, raffinose, fructose, lactulose, sodium chloride,
polyethyleneglycol (PEG), maltodextrin, dextran (e.g. dextran 70),
hydroxyethyl starch, fluid gelatin, a synthetic colloid, an
antibody, a binding protein, and albumin.
95. A ductal access system as in claim 93, wherein the container
comprises a syringe for connection to the first side port.
96. A ductal access system as in claim 93, wherein the premeasured
volume is in the range from 2 ml to 100 ml.
97. A ductal access system as in claim 93, wherein the ductal
access fluid is selected from the group consisting of saline,
phosphate buffered saline, a nonabsorbable fluid, an isotonic
solution, an osmotic solution, a hypotonic solution, and a
hypertonic solution.
98. A ductal access device comprising: a hub having an internal
elongate manifold, a lower port at a bottom of the manifold, and
first and second side ports spaced above the lower port; and an
access tube having a distal end, a proximal end, a lumen
therethrough, and dimensions which permit introduction of the
distal end through a ductal orifice and a positioning a distal end
thereof distal to the ductal sphincter of the human breast, wherein
the proximal end of the tube is attached to the lower port of the
hub.
99. A ductal access device as in claim 98, wherein the first and
second side ports are at the same level relative to the lower
port.
100. A ductal access device as in claim 98, wherein the first side
port is below the second side port.
101. A ductal access device as in claim 98, wherein the access tube
has an outer diameter of 0.010 inches or 0.245 mm or greater.
102. A ductal access device as in claim 98, wherein the access tube
has an outer diameter of 0.50 inches or 1.27 mm or less.
103. A ductal access device as in claim 101, wherein the outer
diameter is in the range from 0.010 inches or 0.245 mm to 0.050
inches or 1.27 mm.
104. A ductal access device as in claim 103, wherein the access
tube has a lumen diameter 0.007 inches (0.178 mm) or greater.
105. A ductal access device as in claim 103, wherein the access
tube has a lumen diameter in a range from 0.007 inches (0.178 mm)
to 0.047 inches (1.19 mm).
106. A ductal access device as in claim 98, further comprising: an
infusion tube connected to the first port of the hub; and a
collection tube connected to the second port of the hub.
107. A ductal access device as in claim 98, wherein the manifold
has a volume in the range from 0.01 cc to 1.0 cc.
108. A ductal access device as in claim 98, wherein the first side
port is spaced above the lower port by a distance less than 5 mm
and the second side port is spaced above the first side port by a
distance in the range from 0.10 mm to 5 mm.
109. A ductal access device as in claim 98, further comprising a
dilator removably received in the hub and having a distal tip which
is positionable through the access tube to extend from the distal
end thereof.
110. A ductal access device as in claim 109, wherein the dilator
has an outer diameter of 0.024 inches (or 0.061 mm) or less.
111. A ductal access device as in claim 109, wherein the dilator is
positionable through the hub manifold and into the lumen of the
access tube.
112. A ductal access device as in claim 109, wherein the dilator is
tapered.
113. A ductal access device as in claim 109, wherein a receiving
portion of the hub for receiving the dilator comprises a
water-tight seal.
114. A ductal access device as in claim 98, further comprising
means on the access tube for positioning the distal end of the
access tube distal to the ductal sphincter.
115. A ductal access device as in claim 114, wherein the
positioning means comprises length indicia on the tube which permit
a user to determine the depth to which the distal end of the tube
has been introduced.
116. A ductal access device as in claim 114, wherein the
positioning means comprises a stop element formed or attached to
the tube, wherein the stop has dimensions which prevent further
insertion of the tube into the duct and wherein the stop is
positioned on the tube so that the distal tip will be located
distal to the ductal sphincter when the device is fully inserted up
to the stop.
117. A ductal access device as in claim 116, wherein the stop
element comprises a collar affixed to or formed on an exterior
surface of the tube.
118. A ductal access device as in claim 116, wherein the stop
element comprises a hub attached to a proximal end of the tube,
wherein the hub has a width which is greater than the diameter of
the tube so that a shoulder is formed at a junction between the
tube and the hub.
119. A ductal access device as in claim 98, further comprising
means for anchoring the device to the breast.
120. A ductal access device as in claim 114, wherein the
positioning means comprises a nob on the access tube having an
increase diameter for anchoring the tube distal to a ductal
sphincter once the nob has passed the sphincter and rests distal to
it.
121. A ductal access system comprising: a ductal access device as
in claim 98; and a container holding a premeasured volume of ductal
wash fluid.
122. A ductal access system as in claim 121, wherein the container
comprises a syringe for connection to the first side port.
123. A ducal access system as in claim 121, wherein the
pre-measured volume is in the range from 2 ml to 100 ml.
124. A ductal access system as in claim 121, wherein the ductal
access fluid is selected from the group consisting of saline,
phosphate buffered saline, a nonabsorbable fluid, an isotonic
solution, an osmotic solution, a hypotonic solution, and a
hypertonic solution.
125. A ductal access system as in claim 121, wherein the ductal
access fluid is selected from the group consisting of a protein, a
colloid, a sugar, a polymer, mannitol, sorbitol, glucose, glycerol,
sucrose, raffinose, fructose, lactulose, sodium chloride,
polyethyleneglycol (PEG), maltodextrin, dextran (e.g. dextran 70),
hydroxyethyl starch, fluid gelatin, a synthetic colloid, an
antibody, a binding protein, and albumin.
126. A ductal access catheter comprising: a catheter body having a
distal end and a proximal end and including at least a distal
portion and a proximal portion; wherein the distal portion has a
cross-sectional geometry which can be inserted through a ductal
orifice into a ductal lumen of a human breast; wherein the proximal
portion has a cross-sectional geometry which inhibits insertion
through the ductal orifice and into the ductal lumen; and wherein
the catheter body has at least an infusion lumen and an collection
lumen each of which has a distal port near a distal end of the
distal portion and a proximal connector near a proximal end of the
proximal portion.
127. A ductal access catheter as in claim 126, wherein the distal
portion of the catheter body is stiffened over at least a part of
its length to facilitate insertion through the ductal orifice and
into the ductal lumen.
128. A ductal access catheter as in claim 127, wherein the
stiffened distal portion of the catheter body has an average
bending stiffness in the range from about 0.010 inch-lbs to about
0.50 inch-lbs.
129. A ductal access catheter as in claim 127, further comprising a
stiffening member disposed in the distal portion of the catheter
body.
130. A ductal access catheter as in claim 126, wherein the distal
portion of the catheter body has a maximum width in the range from
0.008 inches (0.020 mm) to 0.035 inches (0.089 mm) and the proximal
portion of the catheter body has a minimum width in the range from
0.010 inches (0.0254 mm) to 0.100 inches (0.254 mm).
131. A ductal access catheter as in claim 130, wherein the distal
portion of the catheter body has a generally tubular structure with
a diameter in the range from 0.008 inches (0.020 mm) to 0.035
inches (0.089 mm) and the proximal portion of the catheter body has
a generally tubular structure with a diameter in the range from
0.030 inches (0.076 mm) to 0.10 inches (0.254 mm) and wherein the
proximal diameter is greater than the distal diameter by at least
0.010 inches.
132. A ductal access catheter as in claim 126, wherein at least one
of the distal collection port and the distal infusion portion are
disposed on a side of the distal portion of the catheter body.
133. A ductal access catheter as in claim 132, wherein the distal
collection port and the distal infusion port are both located on
the side of the distal portion of the catheter body.
134. A ductal access catheter as in claim 133, wherein the distal
collection port and the distal infusion port are axially
aligned.
135. A ductal access catheter as in claim 133, wherein the distal
collection port and the distal infusion port are axially spaced
apart.
136. A ductal access catheter as in claim 133, wherein the catheter
body includes an atraumatic distal tip.
137. A ductal access catheter as in claim 136, wherein the tip is
composed of a soft polymeric material, has a diameter in the range
from about 0.008 inches (0.020 mm) to about 0.035 inches (0.089
mm), and a length at least 0.25 cm.
138. A ductal access catheter comprising: a catheter body having a
distal end and a proximal end and including at least a distal
portion and a proximal portion; wherein the distal portion has a
cross-sectional geometry which can be inserted through a ductal
orifice into a ductal lumen of a human breast; wherein the distal
portion of the catheter body is stiffened over at least a part of
its length to facilitate insertion through the ductal orifice and
into the ductal lumen; and wherein the catheter body has at least
an infusion lumen and an collection lumen each of which has a
distal port near a distal end of the distal portion and a proximal
connector near a proximal end of the proximal connector.
139. A ductal access catheter as in claim 138, wherein the
stiffened distal portion of the catheter body has an average
bending stiffness in the range from about 0.010 inch-lbs to about
0.50 inch-lbs.
140. A ductal access catheter as in claim 138, wherein the proximal
portion has a cross-sectional geometry which inhibits insertion
through the ductal orifice and into the ductal lumen.
141. A ductal access catheter comprising: a catheter body having a
distal end and a proximal end and including at least a distal
portion and a proximal portion; wherein the distal portion has a
cross-sectional geometry which can be inserted through a ductal
orifice into a ductal lumen of a human breast; and wherein the
catheter body has at least an infusion lumen and an collection
lumen each of which has a distal port near a distal end of the
distal portion and a proximal connector near a proximal end of the
proximal connector; and wherein the distal collection port and the
distal infusion port are both located on the side of the distal
portion of the catheter body.
142. A ductal access catheter as in claim 141, wherein the distal
collection port and the distal infusion port are axially
aligned.
143. A ductal access catheter as in claim 141, wherein the distal
collection port and the distal infusion port are axially spaced
apart.
144. A ductal access catheter as in claim 141, wherein the proximal
portion has a cross-sectional geometry which inhibits insertion
through the ductal orifice and into the ductal lumen.
145. A method for lavage of a ductal network in a human breast,
said method comprising: providing a catheter as in claim 127;
inserting the distal portion of the catheter through a ductal
orifice and into a distal lumen of the ductal network; introducing
a wash fluid through the infusion lumen into the ductal network;
and withdrawing the wash fluid and substances borne by the wash
fluid from the ductal network through the collection lumen.
146. A ductal access system comprising: a catheter as in claim 127,
and instructions for use setting forth a method for lavage of a
ductal network in a human breast including introducing a wash fluid
through the infusion lumen into the ductal network and withdrawing
the wash fluid and substances borne by the wash fluid from the
ductal network through the collection lumen.
147. A device as in claim 52, further comprising a means for
controlling a flow of fluid through the infusion lumen.
148. A device as in claim 52, further comprising a means for
controlling a flow of fluid through the collection lumen.
149. A device as in claim 52, further comprising both a means for
controlling a fluid flow through the infusion lumen and a means for
controlling a fluid flow through the collection lumen.
150. A device as in claim 149, wherein the fluid control means
comprise compressable lumens.
151. A device as in claim 149, wherein the fluid control means
comprise stopcocks on each lumen.
152. A device as in claim 106, further comprising a means for
controlling a flow of fluid through the infusion tube.
153. A device as in claim 106, further comprising a means for
controlling a flow of fluid through the collection tube.
154. A device as in claim 106, further comprising both a means for
controlling a fluid flow through the infusion lumen and a means for
controlling a fluid flow through the collection lumen.
155. A device as in claim 154, wherein the fluid control means
comprise compressable lumens.
156. A device as in claim 154, wherein the fluid control means
comprise stopcocks on each lumen.
157. A method for increasing an amount of fluid collectable from a
milk duct of a breast of a mammal comprising administering an agent
to a ductal lumen of a breast capable of maintaining or increasing
the amount of collectable fluid in the ductal lumen, and collecting
the fluid from the duct.
158. The method of claim 157, wherein the agent comprises an agent
selected from the group consisting of a nonabsorbable agent, an
oncotic agent and an osmotic agent.
159. The method of claim 157, wherein the agent is soluble.
160. The method of claim 157, wherein the agent comprises a
molecule selected from the group consisting of a protein, a
colloid, a sugar, and a polymer.
161. The method of claim 160, wherein the agent comprises a protein
and the protein is selected from the group consisting of a binding
protein and an antibody.
162. The method of claim 161, wherein the protein is a binding
protein, and the binding protein comprises albumin.
163. The method of claim 157, wherein the agent comprises an agent
selected from the group consisting of mannitol, sorbitol, glucose,
glycerol, sucrose, raffinose, fructose, lactulose, sodium chloride,
albumin, polyethyleneglycol (PEG), maltodextrin, dextran (e.g.
dextran 70), hydroxyethyl starch, fluid gelatin, and a synthetic
colloid.
164. The method of claim 157, wherein administering comprises
administering locally.
165. The method of claim 164, wherein administering locally
comprises administering intraductally.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application claims the benefit of each of the following
provisional applications under 37 CFR .sctn.1.78: No. 60/114,048,
filed on Dec. 28, 1998; No. 60/134,613, filed on May 18, 1999; No.
60/143,476, filed on Jul. 12, 1999; No. 60/143,359, filed on Jul.
12, 1999; and No. 60/______ (attorney docket no. 18612-003100),
filed on Dec. 14, 1999. The fill disclosures of each these
applications are incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The field of this invention is devices, methods and systems
for collecting breast duct fluid from humans.
[0004] 2. Description of the Background Art
[0005] For several decades significant members of the medical
community dedicated to studying breast cancer have believed and
shown that the cytological analysis of cells retrieved from nipple
discharge from the breast milk ducts can provide valuable
information leading to a identifying patients at risk for breast
cancer. Indeed Papanicolaou himself contributed to the genesis of
such a possibility of a "Pap" smear for breast cancer by analyzing
the cells contained in nipple discharge. See Papanicolaou et al,
"Exfoliative Cytology of the Human Mammary Gland and Its Value in
the Diagnosis of Cancer and Other Diseases of the Breast" Cancer
(1958) March/April 377-409. See also Petrakis, "Physiological,
biochemical, and cytological aspects of nipple aspirate fluid",
Breast Cancer Research and Treatment 1986; 8:7-19; Petrakis,
"Studies on the epidemiology and natural history of benign breast
disease and breast cancer using nipple aspirate fluid" Cancer
Epidemiology, Biomarkers and Prevention (Jan/Feb 1993) 2:3-10;
Petrakis, "Nipple Aspirate Fluid in epidemiological studies of
breast disease", Epidemiologic Reviews (1993) 15:188-195. More
recently, markers have also been detected in nipple fluid. See
Sauter et al, "Nipple aspirate fluid: a promising non-invasive
method to identify cellular markers of breast cancer risk", British
Journal of Cancer 76(4):494-501 (1997). The detection of CEA in
fluids obtained by a nipple blot is described in Imayama et al.
(1996) Cancer 78: 1229-1234.
[0006] Breast cancer is believed to originate in the lining of a
single breast milk duct in the breast; and additionally human
breasts are believed to contain from 6 to 8 of these ducts. See
Sartorius, JAMA 224 (6): 823-827 (1973). Sartorious describes use
of hair-like single lumen catheters that are inserted into breast
ducts using an operating microscope and the ducts were flushed with
saline solution as described in Cassels, D Mar. 20.sup.th, 1973,
The Medical Post, article entitled "New tests may speed breast
cancer detection". Sartorius et al, Contrast ductography for
recognition and localization of benign and malignant breast
lesions: an improved technique. pp. 281-300. In: Logan WW, ed.
BREAST CARCINOMA New York, Wiley, 1977. After the fluid was
infused, the catheter was removed because it was too small to
collect the fluid, the breast was squeezed and fluid that oozed
onto the nipple surface was removed from the surface by a capillary
tube. Similarly, Love and Barsky, "Breast-duct endoscopy to study
stages of cancerous breast disease", Lancet 348(9033):997-999, 1996
describes cannulating breast ducts with a single lumen catheter and
infusing a small amount of saline, removing the catheter and
squeezing to collect the fluid that returns on the nipple surface.
The use of a rigid 1.2 mm ductscope to identify intraductal
papillomas in women with nipple discharge is described in Makita et
al (1991) Breast Cancer Res Treat 18: 179-188. It would be
advantageous to develop methods and devices to collect the ductal
fluid from within the duct.
[0007] Galactography, or contrast ductography has for years located
breast ducts based on spontaneous nipple discharge, infused the
ducts (using cannulas for this purpose) with contrast dye
solutions, and taken x-ray pictures to determine the source of the
discharge within the duct. See generally, The Breast: Comprehensive
Management of Benign and Malignant Breast Diseases, Bland and
Copeland eds. W. B. Saunders Co. Philadelphia Pa. 1991 pages
61-67.
[0008] Method and kits for obtaining fluid and cellular material
from breast ducts Ser. No. 09/067,661 filed Apr. 28, 1998, and its
CIP Ser. No. 09/301,058 filed Apr. 28, 1999 describe and claim
infusing a small amount of fluid into the duct and collecting the
fluid using a catheter. It would be beneficial to optimize the
cells and fluid collected from this procedure.
[0009] U.S. Ser. No. 60/143,359 filed Jul. 12, 1999 describes and
claims a multilumen catheter for collection of infused fluid. U.S.
Ser. No. 60/143,476 filed Jul. 12, 1999 describes and claims
devices and methods for accessing the lactiferous sinus of a breast
duct. U.S. Ser. No. 60/122,076 filed Mar. 1, 1999 describes
devices, methods and kits for accessing more than one breast duct
at a time for delivering and/or retrieving agents or materials to
and from more than one breast duct at the same time. Related
applications are U.S. Ser. Nos. 60/143,476 and No. 60/143,359 both
filed Jul. 12, 1999 and No. 60/134,613 filed May 18, 1999, and No.
60/114,048 filed Dec. 28, 1998, all of which are herein
incorporated by reference in their entirety.
[0010] Osmotic agents including sugars that are poorly absorbed,
for example lactulose or sorbitol, have been used as laxatives. See
THE MERCK MANUAL OF MEDICAL INFORMATION, Berkow, Beers and Fletcher
Eds, 1997 Merck Res. Lab., Whitehouse Station, N.J. pp. 522-523.
The osmotic agent mannitol is available as an injectable, 25%
(Physicians Desk Reference 1996) for a variety of indications (e.g.
renal insufficiency, congestive heart failure). A mixture of
sorbitol and mannitol is compared to distilled water as an irrigant
during transurethral prostatectomy in Sargin et al, (1997) Int Urol
Nephrol 29:575-80. Intracranial pressure therapy has been provided
by solutions of mannitol, sorbitol or glycerol as described in
Treib et al, (1998) Eur Neurol 40: 212-219. Osmotherapy for
increased intracranial pressure comparing the use of mannitol and
glycerol is discussed in Biestro et al, (1997) Acta Neurochir
(Wien) 138: 725-32; discussion 732-3. Mannitol therapy for renal
conditions is described generally in Better et al, (1997) Kidney
Int 52:886-894, and use of the osmotic diuretic mannitol for renal
protection is analyzed in Visweswaran et al, (1997) J Am Soc
Nephrol 8: 1028-33. Use of mannitol during cardiac catheterization
is described in Willerson et al, (1975) Circulation 51:1095-1100
and Kurnick et al, (1991) Am J Kidney Dis 17: 62-8. The osmotic
effects of polyethylene glycol are discussed in Schiller et al,
(1988) Gastroenterology 94: 933-41. Raffinose is used for
peritoneal dialysis as described in Kohan et al (1998) J Lab Clin
Med 131: 71-6.
[0011] Relevant Literature
[0012] Hou et al, "A simple method of Duct Cannulation and
Localization for Galactography before Excision in Patients with
Nipple Discharge." Radiology 1995; 195; 568-569 describes injecting
a "small volume of sterile, water soluble contrast material . . .
(0.5 ml-2.0 ml) . . . the catheter was taped on the breast or
nipple . . . the contrast material was aspirated with the same
syringe and gentle manual pressure was exerted on the breast to
expel the opaque medium."
[0013] The use of a 0.4 mm flexible scope to investigate nipple
discharge is described in Okazaki et al (1991) Jpn J. Clin. Oncol.
21:188-193 in which before the fiberoptic ductoscopy "a lacrimal
cannula was inserted [into the duct] for ductal washing by infusing
0.2 to 0.5 ml physiological saline twice or three times, citing
also Okazaki et al Nyugan No Ringsho 4:587-594 (1989) (in
Japanese).
[0014] A company called Diagnostics, Inc. formed in 1968, produced
devices to obtain breast ductal fluid for cytological evaluation.
The devices included a hair-like single lumen breast duct catheter
to infuse fluid into a breast duct and the procedure dictated that
after removal of the catheter oozing fluid was collected from the
nipple surface with a capillary tube. The devices were sold prior
to May 28, 1976 for the purpose of collecting breast ductal fluid
for cytological evaluation.
[0015] A lacrimal irrigating cannula is described in U.S. Pat. No.
5,593,393 to inventors Trudell and Prouty. The cannula is graduated
and used for insertion, dilation, probing and irrigating of the
lacrimal drainage system of the eye. Lacrimal probes have been used
to access breast ducts as depicted in The Breast: Comprehensive
Management of Benign and Malignant Diseases (1991) vol 2, Bland
& Kirby eds. W.B. Saunders Co, Philadelphia, Pa. pp. 63, FIG.
3-26.
[0016] Patents and applications that describe use of a fixed
support wire or support generally to reinforce the catheter include
PCT publication WO 97/44084, PCT publication WO 97/44082, U.S. Pat.
No. 5,221,255, JP 6-154334 (unexamined patent publication), U.S.
Pat. No. 3,792,703 to Moorehead, U.S. Pat. No. 4,596,564 to
Spetzler et al, U.S. Pat. No. 5,209,734 to Hurley et al, U.S. Pat.
No. 5,456,674 to Bos et al, PCT publication WO 97/31677, PCT
publication WO 94/07549, PCT publication WO 94/02197, European
patent application EP 630 657 A1, European patent application EP
800 842 A1, Japanese unexamined patent publications JP 4-226675, JP
6-277289, and JP 6-277294, Japanese examined patent publication JP
3-4232, and PCT publication WO 97/47230.
[0017] Patents and publications that describe use of a very small
atraumatic tip include U.S. Pat. No. 4,652,255 to Martinez, U.S.
Pat. No. 5,246,430 to MacFarlane, PCT publication WO 97/37699, PCT
publication WO 97/10015, PCT publication WO 94/07549, European
patent application EP 729 766 A1, European patent application EP
643 979 A1, Japanese examined utility model publication JP 4-4730,
and Japanese examined patent publications JP 1-14794, JP 61-24022,
and JP 61-24023.
[0018] Patents and publications that describe and claim fluid
collection catheters having a narrow distal portion and a larger
diameter proximal portion with a shoulder therebetween include PCT
publication WO 97/44084, PCT publication WO 97/44082, U.S. Pat. No.
5,221,255 to Mahurkar, JP 2,519,873 (U.S. Pat. No. 5,470,318), U.S.
Pat. No. 4,553,957 to Williams et al, U.S. Pat. No. 4,652,255 to
Martinez, U.S. Pat. No. 4,709,705 to Truglio, U.S. Pat. No.
5,451,208 to Goldrath, U.S. Pat. No. 5,246,430 to MacFarlane, PCT
publication WO 97/48435, PCT publication WO 97/31677, PCT
publication WO 95/20983, PCT publication WO 94/02197, European
patent application EP 682 954 A2, European patent application EP
631 791 A1, Japanese examined patent publication JP 4-45186,
Japanese unexamined utility model publication 6-77709, PCT
publication WO 98/39046, and WO 97/47230.
[0019] Other patents or publications related in the art include the
following: JP 5-184664 assigned to Terumo Corp. describes a
catheter with a distal tip formed by heating; JP 2.631,320 Moriuchi
et al, assigned to Terumo Corp. showing vascular catheter with
multiple axial wire supports extending the length of the catheter;
JP 3-264045 to Sato, assignee Terumo Corp. has a central
reinforcement wire extending the length of intravascular catheter
body; JP 61-268266 (WO 89109079) to Hurley et al, assignee Sumitomo
Bakelite (abandoned) depicting another wire reinforcement but in a
uterine catheter; JP 6-502314 to Hurley et al, assignee Brigham
& Women's Hospital shows a spinal catheter with spinal wire
reinforcement; JP 8-112354 to Takane depicts probe with isolate
lumens and distal side ports; JP 5-237191 (EP 542246) to Pearsall,
assignee Becton Dickinson shows rounded tips softer than the body
of the catheter; JP 3-36363 (JP 4-516C) to Kamogawa, assignee
Terumo Corp. is expired but has atraumatic tip with side ports and
a single lumen; JP 2,531,583 to Onishi, assigned to Mitsubishi
shows a catheter having a soft tip formed from polymer having a
glass transition temperature at body temperature; JP 2,681,345 to
Inoue, assignee Kitasato Supply shows insemination device with
syringe; JP 5-184664 to Takeoka, assignee Terumo Corp. shows a
single lumen rounded tip catheter with side ports; JP 58-46337 (JP
59-2345) to Fujimoto depicts a slidable stop on rectal catheter
having side ports; and JP 58-146356 to Harris depicts an
intrauterine catheter with shoulder stop. Patents and publications
that describe breast access for purposes other than lavage include
U.S. Pat. No. 5,800,534 to Jeter et al.
SUMMARY OF THE INVENTION
[0020] According to the present invention, a method for obtaining
cellular material from a human breast milk duct comprises
introducing a wash fluid to the breast milk duct, using a volume of
at least 2 ml that is present within the duct for a preselected
time, and collecting at least a portion of the introduced wash
fluid from within the duct, with the portion of wash fluid carrying
the cellular material. The preselected time is preferably less than
one second, but will usually be in the range from one second to one
hour. The wash fluid is preferably introduced to a volume of at
least 2 ml, often at least 5 ml, and typically in the range between
5 ml and 25 ml, prior to collecting any of the wash fluid from the
duct. The wash fluid is preferably introduced to a single breast
milk duct and collected from the same breast milk duct without
mixing with materials from other breast milk ducts. The method may
further comprise separating cellular material from the collected
fluid. The method may still further comprise examining the
separated cellular material. The cellular material usually includes
a substance selected from the group consisting of whole cells,
cellular debris, proteins, nucleic acids, polypeptides,
glycoproteins, lipids, fats, glycoproteins, small organic
molecules, metabolites, and macromolecules.
[0021] Another aspect of the invention comprises a method for
obtaining cellular material from a human breast milk duct including
introducing a ductal access device having at least one lumen into a
duct, introducing a wash fluid through the access device lumen into
the milk duct, providing a volume of at least 2 ml to be present
within the duct for a preselected time, and then collecting at
least a portion of the wash fluid from the duct through the lumen
of the access device. The method preferably further comprises
massaging and squeezing the breast tissue after introducing the
wash fluid but prior to and/or during collecting a portion of the
wash fluid. Introducing the ductal access device typically
comprises positioning a distal end of the device distal to the
ductal sphincter in the breast duct. The access device preferably
includes only a single lumen that extends into the duct. The wash
fluid is preferably introduced to a volume of at least 2 ml prior
to collecting any of wash fluid from the duct. The preselected time
can be less than one second, but will usually be in the range from
one second to one hour. The wash fluid can be introduced to a
single breast milk duct and collected from the same breast milk
duct without mixing with materials from other breast milk ducts.
The method may still further comprise separating cellular material
from the collected fluid. The method may still further comprise
examining the separated cellular material. The cellular material is
usually a substance selected from the group consisting of whole
cells, cellular debris, proteins, nucleic acids, polypeptides,
glycoproteins, lipids, fats, glycoproteins, small organic
molecules, metabolites, and macromolecules.
[0022] Another aspect of the invention is a method for obtaining
cellular material from a human breast milk duct comprising
introducing a wash fluid to the breast milk duct, providing that
the wash fluid is present within the duct for a preselected time,
and collecting at least a portion of the introduced wash fluid from
within the duct, where the portion carries the cellular material;
the wash fluid is introduced to a single breast milk duct and
collected from the same breast milk duct without mixing with
materials from other breast milk ducts. The volume of wash fluid
can be at least 2 ml. The preselected time can be less than one
second or can be in a range from one second to one hour. The method
can further comprise separating cellular material from the
collected fluid. The method can also further comprise examining the
separated cellular material. The cellular material can be a
substance selected from the group consisting of whole cells,
cellular debris, nucleic acids, lipids, protein metabolites, small
organic molecules, and macromolecules.
[0023] An aspect of the invention is another method for obtaining
cellular material from a human breast milk duct comprising
introducing a ductal access device having at least one lumen into a
duct, introducing a wash fluid through the access device lumen into
the milk duct, where the wash fluid is present within the duct for
a preselected time, and collecting at least a portion of the wash
fluid from the duct through the lumen of the access device; the
wash fluid is introduced to a single breast milk duct and collected
from the same breast milk duct without mixing with materials from
other breast milk ducts. The volume of wash fluid can be at least 2
ml. The preselected time can be less than one second or in a range
from one second to one hour. The method can further comprise
separating cellular material from the collected fluid, and the
separated material can be examined. The cellular material can be a
substance selected from the group consisting of whole cells,
cellular debris, nucleic acids, lipids, protein metabolites, small
organic molecules, and macromolecules.
[0024] An aspect of the invention is a kit comprising a ductal
access device; and instructions for use setting forth a method
provided above comprising introducing a ductal access device having
at least one lumen into a duct.
[0025] An aspect of the invention is a ductal access device
comprising an access tube having a distal end, at least one lumen,
and dimensions which permit introduction of the distal end through
a ductal orifice and positioning a distal end distal to the ductal
sphincter of a human breast. The device can further comprise means
on the access tube for positioning the distal end distal to the
ductal sphincter. The positioning means can comprise length indicia
on the tube which permit a user to determine the depth to which the
distal end of the tube has been introduced. The positioning means
can comprise a stop element formed or attached to the tube; the
stop will have dimensions which prevent further insertion of the
tube into the duct and the stop is positioned on the tube so that
the distal tip will be located distal to the ductal sphincter when
the device is fully inserted up to the stop. The stop element can
comprise a collar affixed to or formed on an exterior surface of
the tube. The device can comprise means for anchoring the device to
the breast. The device can include a receiving portion comprising a
water tight seal for receiving the dilator. The stop element can
comprise a hub attached to a proximal end of the tube, where the
hub has a width which is greater than the diameter of the tube so
that a shoulder is formed at a junction between the tube and the
hub. The positioning means can comprise a nob on the access tube
having an increase diameter for anchoring the tube in the
lactiferous sinus once the nob has passed the sphincter and rests
in the sinus. The access tube can have an outer diameter of 0.05
inches (or 1.27 mm) or less. The access tube can have an outer
diameter of 0.010 inches (or 0.254 mm) or greater. The outer
diameter can be in the range from 0.010 inches (or 0.254 mm) to
0.050 inches (or 1.27 mm). The access tube can have a lumen
diameter 0.007 inches (or 0.178 nm) or greater. The access tube can
have a lumen diameter in the range from 0.007 inches (or 0.178 mm)
to 0.047 inches (or 1.19 mm). The access device can further
comprise an infusion connector providing a fluid flow path into the
lumen of the tube; and a collection connector providing a fluid
outlet path from the lumen of the tube; the infusion and collection
connectors are isolated from each other so that the fluid may be
infused through the infusion connector and simultaneously removed
through the collection connector. The device can further comprise a
dilator removably received in the access tube and having a distal
tip which is positionable through the access tube to extend from
the distal end of the device. The dilator can have an outer
diameter of 0.024 inches (or 0.61 mm) or less. The dilator can be
tapered. A receiving portion of the device for receiving the
dilator can comprise a water-tight seal.
[0026] An aspect of the invention is a ductal access system
comprising a ductal access device as described and a container
holding a premeasured volume of ductal wash fluid. The container
can comprises a syringe for connection to the first side port. The
pre-measured volume can be in the range from 2 ml to 100 ml. The
ductal access fluid is can be selected from the group consisting of
saline, phosphate buffered saline, a nonabsorbable fluid, an
isotonic solution, an osmotic solution, a hypotonic solution, and a
hypertonic solution.
[0027] A further aspect of the invention is a ductal access device
comprising an access tube having a distal end, a single lumen, and
dimensions which permit introduction of the distal end through a
ductal orifice and positioning a distal end of the device distal to
the ductal sphincter, an infusion connector providing a fluid flow
path into the lumen of the access tube, and a collection connector
providing a fluid outlet path from the lumen of the access tube;
the infusion and collection connectors being isolated from each
other so that fluid may be infused through the infusion connector
and simultaneously removed through the collection connector. The
tube has an outer diameter of 0.010 inches (or 0.254 mm) or greater
or the tube has an outer diameter of 0.050 inches (or 1.27 mm) or
less, or the outer diameter can be in the range from 0.010 inches
(or 0.254 mm) to 0.50 inches (or 1.27 mm).
[0028] The access tube has a lumen diameter 0.007 inches (or 0.178
mm) or greater, or a lumen diameter in the range from 0.007 inches
(or 0.178 mm) to 0.047 inches (or 1.19 mm). The device can further
comprise means on the access tube positioning a distal end of the
device distal to the ductal sphincter. The positioning means can
comprise length indicia on the tube which permit a user to
determine the depth to which the distal end of the tube has been
introduced. The positioning means comprises a stop element formed
or attached to the tube, and the stop has dimensions which prevent
further insertion of the tube into the duct; the stop is positioned
on the tube so that a distal end of the distal tip is positioned
distal to the ductal sphincter. The stop element an comprise a
collar affixed to or formed on an exterior surface of the tube. The
stop element can comprise a hub attached to a proximal end of the
tube, where the hub has a width which is greater than the diameter
of the tube so that a shoulder is formed at a junction between the
tube and the hub. The positioning means can comprise a nob on the
access tube having an increased diameter for anchoring the distal
portion of the tube distal to the sphincter once the nob has passed
the sphincter. The device can comprise means for anchoring the
device to the breast. The device can additionally comprise a
dilator removably received in the access tube and having a distal
tip which is positionable through the access tube to extend from
the distal end of access device. The dilator can have an outer
diameter of 0.024 inches (or 0.61 mm) or less. The dilator can be
tapered. A receiving portion of the device for receiving the
dilator an comprise a water-tight seal.
[0029] An aspect of the invention is a ductal access system
comprising a ductal access device as just described and a container
holding a premeasured volume of ductal wash fluid. The container
can comprise a syringe for connection to the first side port. The
premeasured volume can be in the range from 2 ml to 100 ml. The
ductal access fluid can be selected from the group consisting of
saline, phosphate buffered saline, a nonabsorbable fluid, an
isotonic solution, an osmotic solution, a hypotonic solution, and a
hypertonic solution.
[0030] An aspect of the invention is a ductal access device
comprising a hub having an internal elongate manifold, a lower port
at a bottom of the manifold, and first and second side ports spaced
above the lower port; and an access tube having a distal end, a
proximal end, a lumen, and dimensions which permit introduction of
the distal end through a ductal orifice and a positioning a distal
end of the device distal to the ductal sphincter of the human
breast, provided also that the proximal end of the tube is attached
to the lower port of the hub. The first and second side ports can
be at the same level relative to the lower port. The first side
port can be below the second side port. The access tube can have an
outer diameter of 0.010 inches (or 0.254 mm) or greater. The access
tube can have an outer diameter of 0.50 inches (or 1.27 mm) or
less. The outer diameter can be in the range from 0.010 inches (or
0.245 mm) to 0.50 inches (or 1.27 mm). The access tube can have a
lumen diameter 0.007 inches (0.178 mm) or greater, or a lumen
diameter in a range from 0.007 inches (0.178 mm) to 0.047 inches
(1.19 mm). The device can have an infusion tube connected to the
first port of the hub; and a collection tube connected to the
second port of the hub. The device can further comprise a means for
controlling a flow of fluid through the infusion tube, a means for
controlling a flow of fluid through the collection tube, or both a
means for controlling a fluid flow through the infusion lumen and a
means for controlling a fluid flow through the collection lumen.
The fluid control means can comprise compressable lumens or the
fluid control means can comprise stopcocks on each lumen. The hub
or manifold can have a volume in the range from 0.01 ml to 1.0 ml.
The first side port can be spaced above the lower port by a
distance less than 5 mm and the second side port can be spaced
above the first side port by a distance in the range from 0.10 mm
to 5 mm. The device can further comprise a dilator removably
received in the hub and having a distal tip which is positionable
through the access tube to extend from the distal end of the
device. The dilator can have an outer diameter of 0.024 inches (or
0.61 mm) or less. The dilator can be positionable through the hub
manifold and into the lumen of the access tube. The dilator can be
tapered. A receiving portion of the hub for receiving the dilator
can comprise a water-tight seal. The device can further comprise a
means on the access tube for positioning the distal end of the
access tube distal to the ductal sphincter. The positioning means
can comprise length indicia on the tube which permit a user to
determine the depth to which the distal end of the tube has been
introduced. The positioning means can comprise a stop element
formed or attached to the tube; the stop has dimensions which
prevent further insertion of the tube into the duct and the stop is
positioned on the tube so that the distal tip will be located
distal to the ductal sphincter when the device is fully inserted up
to the stop. The stop element can comprises a collar affixed to or
formed on an exterior surface of the tube. The stop element can
comprise a hub attached to a proximal end of the tube, where the
hub has a width which is greater than the diameter of the tube so
that a shoulder is formed at a junction between the tube and the
hub. The device can further comprise a means for anchoring the
device to the breast. The positioning means can comprise a nob on
the access tube having an increased diameter for anchoring the tube
distal to the ductal sphincter once the nob has passed the
sphincter and rests distal to it.
[0031] An aspect of the invention is a ductal access system
comprising a ductal access device as just described and a container
holding a premeasured volume of ductal wash fluid. The container
can comprise a syringe for connection to the first side port. The
pre-measured volume can be in the range from 2 ml to 100 ml. The
ductal access fluid can be selected from the group consisting of
saline, phosphate buffered saline, a nonabsorbable fluid, an
isotonic solution, an osmotic solution, a hypotonic solution, and a
hypertonic solution.
[0032] An aspect of the invention provides a ductal access catheter
comprising a catheter body having a distal end and a proximal end
and including at least a distal portion and a proximal portion
wherein the distal portion has a cross-sectional geometry which can
be inserted through a ductal orifice into a ductal lumen of a human
breast; wherein the proximal portion has a cross-sectional geometry
which inhibits insertion through the ductal orifice and into the
ductal lumen; and wherein the catheter body has at least an
infusion lumen and an collection lumen each of which has a distal
port near a distal end of the distal portion and a proximal
connector near a proximal end of the proximal portion. The device
can further comprise a means for controlling a flow of fluid
through the infusion lumen, a means for controlling a flow of fluid
through the collection lumen, or both a means for controlling a
fluid flow through the infusion lumen and a means for controlling a
fluid flow through the collection lumen. The fluid control means
can comprise compressable lumens, or the fluid control means can
comprise stopcocks on each lumen
[0033] The distal portion of the catheter body can be stiffened
over at least a part of its length to facilitate insertion through
the ductal orifice and into the ductal lumen. The stiffened distal
portion of the catheter body has an average bending stiffness in
the range from about 0.010 inch-lbs to about 0.5 inch-lbs. The
stiffening member is disposed in the distal portion of the catheter
body.
[0034] The distal portion of the catheter body has a maximum width
in the range from 0.016 inches to 0.022 inches (0.56 mm) and the
proximal portion of the catheter body has a minimum width in the
range from 0.023 inches (0.58 mm) to 0.028 inches (0.71 mm). The
distal portion of the catheter body has a generally tubular
structure with a diameter in the range from 0.010 inches (0.254 mm)
to 0.020 inches (0.51 mm) and the proximal portion of the catheter
body has a generally tubular structure with a diameter in the range
from 0.030 inches (0.762 mm) to 0.10 inches (0.254 mm) and wherein
the proximal diameter is greater than the distal diameter by at
least 0.010 inches (or 0.254 mm). At least one of the distal
collection port and the distal infusion portion can be disposed on
a side of the distal portion of the catheter body. The distal
collection port and the distal infusion port can both be located on
the side of the distal portion of the catheter body. The distal
collection port and the distal infusion port can be axially
aligned. The distal collection port and the distal infusion port
can be axially spaced apart. The catheter body can include an
atraumatic distal tip. The tip can be composed of a soft polymeric
material, have a diameter in the range from about 0.008 inches
(0.20 mm) to about 0.035 inches (0.89 mm), and a length in the
range from about 0.25 cm to about to 2.5 cm.
[0035] The invention further provides a ductal access catheter
comprising a catheter body having a distal end and a proximal end
and including at least a distal portion and a proximal portion;
wherein the distal portion has a cross-sectional geometry which can
be inserted through a ductal orifice into a ductal lumen of a human
breast; wherein the distal portion of the catheter body is
stiffened over at least a part of its length to facilitate
insertion through the ductal orifice and into the ductal lumen; and
wherein the catheter body has at least an infusion lumen and an
collection lumen each of which has a distal port near a distal end
of the distal portion and a proximal connector near a proximal end
of the proximal connector. The stiffened distal portion of the
catheter body can have an average bending stiffness in the range
from about 0.010 inch-lbs to about 0.5 inch-lbs. The proximal
portion can have a cross-sectional geometry that inhibits insertion
through the ductal orifice and into the ductal lumen.
[0036] The invention also provides a ductal access catheter
comprising a catheter body having a distal end and a proximal end
and including at least a distal portion and a proximal portion;
wherein the distal portion has a cross-sectional geometry which can
be inserted through a ductal orifice into a ductal lumen of a human
breast; and wherein the catheter body has at least an infusion
lumen and an collection lumen each of which has a distal port near
a distal end of the distal portion and a proximal connector near a
proximal end of the proximal connector; and wherein the distal
collection port and the distal infusion port are both located on
the side of the distal portion of the catheter body. The distal
collection port and the distal infusion port can be axially
aligned. The distal collection port and the distal infusion port
can be axially spaced apart. The proximal portion can have a
cross-sectional geometry that inhibits insertion through the ductal
orifice and into the ductal lumen.
[0037] Another aspect of the invention is a method for lavage of a
ductal network in a human breast comprising providing a multi-lumen
catheter as just described and inserting the distal portion of the
catheter through a ductal orifice and into a distal lumen of the
ductal network; introducing a wash fluid through the infusion lumen
into the ductal network; and withdrawing the wash fluid and
substances borne by the wash fluid from the ductal network through
the collection lumen.
[0038] Another aspect of the invention is a system comprising a
multi-lumen catheter as just described and instructions for use
setting forth a method for lavage of a ductal network in a human
breast including introducing a wash fluid through the infusion
lumen into the ductal network and withdrawing the wash fluid and
substances borne by the wash fluid from the ductal network through
the collection lumen.
[0039] The agent infused into the duct can comprise a
non-absorbable fluid and/or an oncotic agent and/or an osmotic
agent. The agent can be soluble. The agent can comprise a molecule
that is a protein, a colloid, a sugar, or a polymer. The agent can
be mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, polyethyleneglycol (PEG),
maltodextrin, dextran (e.g. dextran 70), hydroxyethyl starch, fluid
gelatin, or a synthetic colloid. The agent can comprise a protein
and the protein can be a binding protein or an antibody. The
binding protein can be albumin. Administering can comprise
administering locally, and local administration can comprise
administering intraductally. A system for increasing or
standardizing an amount of fluid collectable from a milk duct of a
breast can comprise infusing a nonabsorbable fluid and/or an
osmotic agent and/or an oncotic agent into the ductal lumen, a
medical tool for delivering the agent to the ductal lumen, and
instructions for use.
BRIEF DESCRIPTION OF THE DRAWINGS
[0040] FIG. 1 shows a single lumen catheter with a stop and
external infusion and collection tubes.
[0041] FIG. 2 is a detailed view of a calibrated ductal access
portion of a single lumen catheter. The calibration serves to
identify a depth of penetration.
[0042] FIG. 3 is a single lumen ductal access catheter having a hub
and infusion and collection lumens and a retractable dilator.
[0043] FIG. 3A is a cross section of the device in FIG. 3.
[0044] FIG. 4A illustrates access of a breast duct and penetration
to at least a region distal to the ductal sphincter.
[0045] FIG. 4B illustrates filling a duct with infusion fluid.
[0046] FIG. 4C illustrates bidirectional flow of infused fluid in
the duct through the access lumen to be collected.
[0047] FIG. 4D illustrates a single lumen catheter accessing a
breast duct having the capacity to infuse and collect fluid outside
the accessed breast duct.
[0048] FIG. 5 depicts a kit comprising a single lumen catheter
having infusion and collection lumens outside the ductal access
portion of the catheter, a premeasured solution to infuse into the
duct and instructions for use of the catheter and wash fluid to
access a breast duct and retrieve cellular material.
[0049] FIG. 6 depicts a single or double lumen catheter having an
infusion and collection lumen outside the catheter with stopcocks
on each external lumen to control fluid flow into or out of each
lumen.
[0050] FIG. 7 illustrates a breast duct accessed by a single or
double lumen catheter having separate infusion and collection
lumens outside the ductal access portion of the catheter, having
also stop cocks on each external lumen for controlling fluid flow
in the lumen, and having an infusion receptacle on the infusion
lumen and a collection receptacle on the collection lumen.
[0051] FIG. 8 illustrates an alternative embodiment of the breast
duct access device of the present invention.
[0052] FIGS. 8A and 8B are cross sectional views taken along lines
8A-8A and 8B-8B of FIG. 8, respectively.
[0053] FIG. 8C is a detailed view of the distal end of the device
of FIG. 8.
[0054] FIGS. 9A and 9B depict alternative transition zones in a
ductal access catheter.
[0055] FIG. 9A is a stepped transition zone.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS OF THE INVENTION
[0056] The following preferred embodiments and examples are offered
by way of illustration and not by way of limitation.
[0057] The invention provides methods for obtaining cellular
material from a human breast duct. A wash fluid is introduced and a
volume of at least 2 ml is allowed to remain in the duct for a
preselected time that can range from less than or about one second
to about an hour, including any length of time in between. During
the time that the wash fluid remains in the breast duct, it may mix
with the ductal fluid already present in the duct, and it may
accumulate cellular material either from the ductal lumen walls or
that already present in the existing resident ductal fluid. The
breast duct may be filled with wash fluid before the wash fluid
mixed with ductal fluid and comprising cellular material is
collected. For example, a wash fluid may be infused into the duct
until a point of resistance to infusion, a which point it may be
considered that the breast duct may is filled with wash fluid, and
the just infused fluid can be allowed to reside in the duct for a
preselected time. Once the time has elapsed, the infused fluid and
the contents of the duct with which it has mixed is collected. If a
ductal access tool is used to access the duct and infuse the fluid
into the duct, the in-dwelling tool can obtain or collect the
infused fluid either through the same lumen that was used to infuse
the wash fluid into the duct originally, or through a separate
second lumen adjacent or coaxial to the infusion lumen. In any
event the access tool remains in place in the duct during the
infusion, filing, preselected waiting time (e.g. less than one
second or about one second to one hour), and collection of the wash
fluid mixed with ductal fluid and cellular material from the breast
duct.
[0058] Methods of the invention include accessing a single breast
duct and obtaining cellular material from that duct without
allowing the cellular material or ductal fluid from the accessed
duct to contact the cellular material or ductal fluid of any other
duct, or cellular material or ductal fluid that happens to be
residing on the nipple surface. Thus is provided the opportunity to
analyze a single individual breast duct separate from other breast
ducts of the patient. The wash fluid can be introduced into the
duct by accessing the breast duct with a ductal access device
having at least one lumen. Infusion of wash fluid into the duct is
provided through the lumen accessing the duct. Collection of the
wash fluid mixed with ductal fluid and comprising cellular material
can also be provided through the lumen accessing the duct. Access
of a single breast duct provides also the opportunity to collect
ductal fluid and cellular material from the accessed breast duct
separate from other breast ducts on the breast, without mixing or
contacting the collected fluids and cellular material with that of
the other ducts, and so providing the opportunity to analyze the
condition of the accessed duct separately.
[0059] During the procedure the breast may be massaged and
squeezed. Massaging and squeezing the breast may facilitate
collection of the infused fluid and mixed ductal fluid and cellular
material. The actions of massaging and squeezing the breast may
also provide some disruption of the cells on the lumen walls,
thereby increasing a yield of cellular material from the procedure.
Collection from a collection lumen (either the same lumen as was
used to infuse or a separate lumen) can be further facilitated in
some cases with aspiration applied into the lumen. Preferably,
where an indwelling tool is used, a single lumen accesses the
breast duct, and external to the breast and breast duct the tool
branches into an infusion lumen and a collection lumen. From this
collection lumen, during the period when the fluid is being
collected from the duct, aspiration may be applied.
[0060] Additionally, where a manifold hub is present in the design
of the access tool, once the wash fluid mixed with ductal fluid and
cellular material is passed out of the duct and into the hub,
collection may be facilitated from the collection lumen without
risk of collapsing the ductal wall, but providing an aspiration
pressure in the collection lumen (e.g. using a syringe and pulling
back to collect material into the syringe). Additionally, or
alternatively, the hub filled with collected material may be
flushed into the collection lumen using an infusion of wash fluid
from the infusion lumen. The fluid flow into and out of the
infusion and collection lumens may be facilitated with means on the
device lumens to stop or open the fluid flow into or out of the
lumens.
[0061] Additionally, when a ductal access device is used to access
a breast duct, the distal end of the device comprising an infusion
and/or collection port or ports is placed distal to the ductal
sphincter to provide an optimal position for infusion and
collection of fluid and/or other agents or materials to and from
the breast duct. Means to assure placement of the distal tip of the
device distal to the ductal sphincter can be provided on the device
as further discussed below in the ductal access device design.
[0062] The wash fluid that is introduced into the duct can comprise
any biocompatable agent or solution. Thus, the wash fluid can
comprise e.g. saline, phosphate buffered saline. Additionally or
alternatively, the wash fluid can comprise an agent or agents or
solution that reduces the ability of the fluid or agent to diffuse
through the ductal wall or otherwise leave the duct and enter other
parts of the body. Accordingly, the wash fluid may comprise a
nonabsorbable fluid, an isotonic solution, an osmotic solution, a
hypotonic solution or a hypertonic solution. Fluid or agents may be
administered to the breast duct in order to facilitate, increase,
and/or optimize the amount of material obtained or obtainable from
the breast duct during the procedure. Agents or solutions that may
comprise the infused wash fluid can include, e.g. protein, colloid,
sugar, polymer, mannitol, sorbitol, glucose, glycerol, sucrose,
raffinose, fructose, lactulose, sodium chloride, polyethyleneglycol
(PEG), maltodextrin, dextran (e.g. dextran 70), hydroxyethyl
starch, fluid gelatin, albumin, a synthetic colloid, an antibody or
part of an antibody, or a binding protein.
[0063] Once the wash fluid had been infused in the duct and the
wash fluid and ductal fluid is collected from a breast duct, the
cellular material can be separated and can be examined. The
cellular material can include, e.g. substances selected from the
group consisting of whole cells, cellular debris, proteins, nucleic
acids, polypeptides, glycoproteins, lipids, fats, glycoproteins,
small organic molecules, metabolites, and macromolecules. Whole
cells can be examined by cytology, or any other suitable method for
analyzing the condition of the cells. Other markers present in the
cellular material, ductal fluid generally, or other material
obtained from the breast duct can be analyzed as is appropriate for
the marker being sought, including e.g. binding assays,
immunohistochemistry, or using other analytical technology for
distinguishing and identifying biological molecules obtained from
biological material.
[0064] Chromosomal abnormalities in ductal epithelial cells can
also provide information and act as a marker to identify cancer or
precancer as described in Mark et al (1999) Cancer Genet Cytogenet
108:26-31; Lundlin and Mertens (1998) Breast Cancer Res Treat
51:1-15; Newsham (1998) Am J Pathol 153:5-9; Larson et al (1998) Am
J Pathol 152:1591-8; Adelaide et al (1998) Genes Chromosomes Cancer
22:186-99; Fejzo et al (1998) Gene Chromosome Cancer 22:105-113;
Dietrich et al (1998) Hum Pathol 12: 1379-82; Cavalli et al (1997)
Hereditas 126:261-8; Adeyinka et al (1997) Cancer Genet Cytogenet
97:119-21; Afify and Mark (1997) Cancer Genet Cytogenet 97:101-5;
Brenner and Aldaz (1997) Prog Clin Biol Res 396: 63-82; Mark et al
(1997) Ann Clin Lab Sci 27:47-56; and Fabian et al 1993 J Cellular
Biochemistry 17G:153-16.
[0065] In addition, exemplary markers are described in Masood S.,
(Prediction of recurrence for advanced breast cancer. Traditional
and contemporary pathologic and molecular markers) Surgical
Oncology Clinics of North America. 4(4):601-32, 1995;
Lopez-Guerrero et al (1999) J Hematother 8(1):53-61; Marjumdar and
Diamandis (1999) Br J Cancer 79(9-10):1594-602; Balleine et al
(1999) Br J Cancer 79 (9-10):1564-71; Houston et al (1999) Br J
Cancer 79(7-8):1220-6; Nikolic-Vukosavljevic et al (1998) Tumori
84(6):691-4; Maguire et al (1998) Int J Biol Markers 13(3):139-44;
Stearns et al (1998) Breast Cancer Res Treat 52(1-3):239-59;
Eiriksdottir et al (1998) Eur J Cancer 34(13):2076-81, and U.S.
Pat. No. 5,169,774. Many known breast cancer markers are discussed
and described in readily available medical text books on breast
cancer. In addition, several markers can be identified and analyzed
in the same sample, e.g. Fabian et al 1993 J. Cellular Biochemistry
17G: 153-16 and Fabian et al 1994 Breast Cancer Res Treat
30(3):263-74 looking at estrogen receptor (ER), epidermal growth
factor receptor (EGFR), mutant p53, HER-2 neu by
immunohistochemistry and aneuploidy by image analysis in fine
needle aspirates.
[0066] Cytological assays that can be performed on the cells
retrieved from a duct or from nipple aspirate can include e.g.
assays described in King et al, J. Nat'l Cancer Inst(1983)
71:1115-21,Wrensch et al. (1992) Am. J. Epidem. 135: 130-141,
Papanicolaou et al, (1958) Cancer, 11:377-409 and Goodson W H &
King E B, Chapter 4: Discharges and Secretions of the Nipple, THE
BREAST: COMPREHENSIVE MANAGEMENT OF BENIGN AND MALIGNANT DISEASES
(1998) 2.sup.nd Ed. vol 2, Bland & Kirby eds. W. B. Saunders
Co, Philadelphia, Pa. pp. 51-74. For example, as described in
Goodson and King (page 60) atypical hyperplasia presents as having
cellular abnormalities, increased coarseness of the chromatin, and
tendency for more single cells as well as groups of cells. With
regard to carcinoma in situ, Papanicolaou et al, described cellular
abnormalities, e.g. nuclear abnormalities diagnosed by cytology of
fluid from nipple secretions containing ductal cells. The cytology
of abnormal cells can also be conducted as described in Sartorius
et al (1977) J. Natl Cancer Inst 59: 1073-1080 and King et al,
(1983) JNCI 71(6) 1115-1121. Atypia and carcinoma in situ are
widely characterized pathologically, as described in Page et al,
(1998) Mod Pathol 11(2): 120-8. The ductal fluid can be analyzed by
cytological techniques by placing some of the fluid on a slide with
a standard cytological stain using a light microscope. The cells
can be studied for atypical growth patterns in individual cells and
clusters of cells using published methods, including Mouriquand J,
(1993) S Karger Pub, "Diagnosis of Non-Palpable Breast Lesions:
Ultrasonographically Controlled Fine-Needle Aspiration: Diagnostic
and Prognostic Implications of Cytology" (ISBN 3805557477); Kline T
S and I K, Pub Igaku-Shoin Medical ""Breast: Guides to Clinical
Aspiration Biopsy" (LSBN 0896401596; Masood, American Society of
Clinical Pathology: November 199S, "Cytopathology of the Breast"
ISBN 0891893806; and Feldman P S, American Society of Clinical
Pathology, November 1984,"Fine Needle Aspiration Cytology and Its
Clinical Applications: Breast and Lung" ISBN 0891891846.
[0067] Other references that discuss cytological analysis and which
give guidance to an analysis of ductal epithelial cells derived
from ductal fluid include Silverman et al, (Can FNA biopsy separate
atypical hyperplasia, carcinoma in situ, and invasive carcinoma of
the breast?: Cytomorphologic criteria and limitations in diagnosis,
Diagnostic Cytopathology) 9(6):713-28, 1993; Masood et al,
(Immunohistochemical differentiation of atypical hyperplasia vs.
carcinoma in situ of the breast) Cancer Detection & Prevention.
16(4):225-35, 1992; Masood et al, (Cytologic differentiation
between proliferative and nonproliferative breast disease in
mammographically guided fine-needle aspirates) Diagnostic
Cytopathology 0.7(6):581-90, 1991; Masood S., (Occult breast
lesions and aspiration biopsy: a new challenge) Diagnostic
Cytopathology. 9(6):613-4, 1993; Masood S., (Prognostic factors in
breast cancer: use of cytologic preparations) Diagnostic
Cytopathology. 13(5):388-95, 1995; Novak and Masood, (Nuclear
grooves in fine-needle aspiration biopsies of breast lesions: do
they have any significance?) Diagnostic Cytopathology. 18(5):333-7,
1998; Sidawy et al, (Interobserver variability in the
classification of proliferative breast lesions by fine-needle
aspiration: results of the Papanicolaou Society of Cytopathology
Study) Diagnostic Cytopathology. 18(2):150-65, 1998; Masood et al,
(Automation in cytology: a survey conducted by the New Technology
Task Force, Papanicolaou Society of Cytopathology) Diagnostic
Cytopathology. 18(l):47-55, 1998; and Frykberg and Masood Copeland
EM 3d. Bland KI., (Ductal carcinoma in situ of the breast) Surgery,
Gynecology & Obstetrics 177(4):425-40, 1993.
[0068] Appropriate animal models for breast cancer therapies have
been described, e.g. McKenzie and Sukumar, (Molecular mechanisms of
chemical carcinogenesis in rodent models) Cancer Treatment &
Research 71:313-29, 1994; Chen et al, (Midkine in the progression
of rat N-nitroso-N-methylurea-induced mammary tumors) Molecular
Carcinogenesis. 17(3):112-6, 1996; and Sukumar et al, (Animal
models for breast cancer) Mutation Research 333(1-2):37-44,
1995.
[0069] In addition to some markers discussed and/or articles or
books cited on breast cancer and breast precancer markers, the
following cancer markers are listed here as exemplary and may be
used as well as other markers to analyze the condition of a breast
duct. Standard assay procedures for identifying the markers can be
used, including antibodies or other binding partners, labels,
stains, pattern analysis (for cells and cell components), and in
general any other chemical or visual identification techniques. The
following are exemplary potential markers for such identification
and analysis: cathepsins (including cathepsin D); maspin, fas, fas
ligand, tissue inhibitor of matrix metalloproteinas-1 (TIMP-1);
chemokines (both C-C and C-X-C type chemokines); collagenases,
metalloproteinases, TIMP's, cathepsins, disrupted basement membrane
epitopes, stromolysin-3; cytokeratins (e.g. keratin 14, B1, KA1,
KA4 and 312C8-1); estrogen and progesterone receptors (or any
androgen or other steroid receptor); growth factor receptors for
members of the fibroblast growth family (FGF) including FGF1-18,
vascular endothelial growth factor (VEGF), insulin-like growth
factor-1 (IGF-I), IGF-II, platelet-derived growth factor (PDGF),
keratinocyte growth factor (KGF), and epithelial growth factor
(EGF); placental growth factor (PLGF), hepatocyte growth factor
(HGF), tumor necrosis factor (TNF), transforming growth factor
(TGF) both alpha and beta forms, and angiopoietin, for example;
growth factors and cytokines including e.g. FGF1-18, VEGF, IGF-I,
IGF-II, PDGF, KGF, EGF, PLGF, HGF, TNF, TGF alpha and beta,
angiopoietin; heat shock proteins (HSP) (e.g. HSP27) 27 (HSP27);
ErB type 1 tyrosine kinase receptors (e.g. Her2 (an EGF receptor)
or any ligand or receptor of the ErbB family of ligands and
receptors); integrins, selectins, cadherins, for example (i.e.
alpha and beta 3 integrin); keratin-14; known cancer antigens
including, for example Ki-67, Ki-S1, p53, nm23, bcl-2, p21 ras,
cyclins, and pS2; thrombin receptor activating peptide; urokinase,
urokinase-type plasminogen activator (UPA), plasmin antiplasmin;
UPA receptor (UPAR), fibrinogen, plasmin activator inhibitor-1 and
2 (PAI-1 and 2); telomerase; antibodies to tumor associated
antigen-72 (TAG-72) (e.g. B72.3, B6.2, and TKH2); carcinoembryonic
antigen (CEA) (see e.g. EP 319,686); prostate specific antigen
(PSA); gross cystic disease fluid protein-15 (GCDFP-15); lactose
dehydrogenase (LDH); chromosomal abnormalities (e.g. aneuploidy or
other abnormalities); S1 protein; alkaline phosphatase; myosin;
sialyl Tn (STn) glycopeptide (e.g. TAG-72); Tn glycopeptide; and
nuclear matrix proteins (as described in provisional patent
application filed 11-17-99 docket no. PDH 99-029, herein
incorporated by reference in its entirety).
[0070] In general, markers can be categorized nonexclusively, and
often in overlapping categories as follows: 1. Markers that are
detected or detectable by virtue of protein expression or
overexpression (detection may occur, e.g. by immunohistochernistry
or in situ hybridization); 2. Markers that are detected or
detectable by virtue of MRNA expression or overexpression
(detection may occur, e.g. by differential display techniques); 3.
Markers that are detected or detectable by virtue of a post
translational change in a protein, e.g. a phosphorylation of the
protein, a ubiquitination, a farnesylation, methylation, or other
modification to the protein that can be detected, e.g. by
antibodies specific to the post translational modification.
[0071] Accordingly, markers such as the following can sought in
ductal fluid, e.g. proteins that are overexpressed, MRNA
transcripts that are over expressed, and proteins comprising post
translational modifications. For example, the following markers can
be identified to distinguish a cancer or precancer cell from a
normal cell. Proteins that are overexpressed can include e.g.
Stromelysin-3, Membrane Type 1 Matrix Metalloproteinase (MT 1-MMP),
Matrix Metalloproteinase-3 (MMP-3), Placental Isoferrintin (p43),
Nuclear Matrix Protein (NMP22), NM-200.4 specific antigen, Vascular
Endothelial Growth Factor (VEGF), Endoglin (CD105), Telomerase,
ErbB-2, ErbB-3, Carcinoembryonic Antigen (CEA), Heat Shock
protein-27 (HSP-27), Breast Cancer-specific Gene (BCSG),
Plasminogen Activator Inhibitor (PAI-1), Urokinase Plasminogene
Activator (uPA), Urokinase Plasminogene Activator Receptor (uPAR),
Colony Stimulating Factor-1 (CSF-1), Colony Stimulating Factor-1
receptor (fins), Annexin I, Vasopressin, the CC Chemokine Regulated
on Activation Normal T cell Expressed and Secreted (RANTES), 44-3A6
specific antigen, A-80 specific antigen, MUC-1, H23 specific
antigen, 83 D4 specific antigen, SP-2 specific antigen, 323/A3
specific antigen, tumor associated antigen-72 (TAG-72), and MBE6
specific antigen.
[0072] Other breast cancer markers detected by any means including
e.g. protein expression, mRNA expression, or post translational
modification can include e.g. (listed alphabetically) alanine
aminopeptidase, alpha 6 integrin, alpha-lactalbumin, AN43, p53,
Bc12-antagonist of cell death (Bad), Bc12-associated athanogene
(BAG-I), Bc12-antagonist/killer 1 (Bak), Bc12-associated X protein
(Bax), Breast cancer antigen 225 (BCA225), B-cell CLL/lymphoma 2
(Bcl-2), Bc12-like 1 (Bcl-x), beta 1-6 branched oligosaccharides,
beta-2 microglobulin (BMG), Bc12 related protein A1 (Bfl-1), bone
sialoprotein (BSP), CCAAT/enhancer-binding protein liver-enriched
inhibitory protein (C/EBPbeta-LIP), Carcinoma Antigen 1 (Ca 1),
Carcinoma Antigen 27.29 (CA 27.29), Carcinoma Antigen M26 (CA M26),
Carcinoma Antigen M29 (CA M29), Carcinoma Antigen 125 (CA125),
Carcinoma Antigen 15.3 (CA15.3), Carcinoma Antigen 195 (CA195),
Carcinoma Antigen 19-9 (CA19-9), Carcinoma Antigen 50 (CA50),
Carcinoma Antigen 549 (CA549), Cadherin-11, calcitonin receptor
(CTR), cathepsin B, cathepsin L, Endoglin (CD105), CD24, CD34
(pan-endothelial marker), CD44, c-met/hepatocyte growth factor
receptor, c-myc, cyclooxygenase-1 (Cox-1), cyclooxygenase-2
(Cox-2), caspase-3 (CPP32), Cyclic nucleotide phosphodiesterase,
cycline E, DNA topoisomerase II-alpha, DNA topoisomerase II-beta,
EGF, EGF receptor, E-selectin, fast homoarginine-sensitive alkaline
phosphatase (FHAP), fatty acid synthase, ferritin, gross cystic
disease fluid protein (GCDFP-15/BRST-2), metastasis-associated
h-mtsl (S 100A4), heat shock cognate protein-73 (hsc73), heat shock
protein-70 (hsp70), heat shock protein-90 alpha (hsp90alpha), heat
shock protein-90 beta (hsp90beta), inhibitors of
differentiation-1(ID1), inhibitors of differentiation-3 (ID3),
interleukin-1 beta, Keratin 8, Keratin 18, Keratin 19, Laminin,
Laminin receptor (MLuC5), Leucine Aminopeptidase (LAP), lipid-bound
sialic acid (LSA), Melanoma antigen-1 (MAGE-1), Melanoma antigen-2
(MAGE-2), Melanoma antigen-3 (MAGE-3), Man6-P glycoproteins,
Mucin-like carcinoma associated antigen (MCA), myeloid cell
leukemia-1 (Mcl-1), metallothionein (MT), mitogen-activated protein
kinase phosphatase-1 (MKP-1), Matrix Metalloproteinase-2 (MMP-2),
Matrix Metalloproteinase-9 (MMP-9), mammary serum antigen (MSA),
breast cancer mucin-2 (MUC-2), breast cancer mucin-3 (MUC-3),
breast cancer mucin-6 (MUC-6), Nm23 nucleoside diphosphate kinase,
omithine decarboxylase (ODC), osteopontin (OPN), P114 (MAR binding
protein), P120 (a nucleolar proliferation antigen), focal adhesion
kinase p125FAK, nuclear autoantigen p330d/CENP-F, plasminogen
activator inhibitor-2 (PAI-2), Pepsinogen C, placental alkaline
phosphatase (PLAP), Platelet factor 4 (angiogenic marker), protein
kinase C (PKC), prostate specific antigen (PSA), pyrimidine
nucleoside phosphorylase, ras p21, reduced glutathione (GSH),
retinoid X receptor alpha, ribosomal S2 protein, sialyltransferase,
Stromelysin-1 (MMP-3), surfactant proteins A, surfactant proteins
B, tumor associated antigen-12 (TAG-12), trefoil gene TFF1, trefoil
gene TFF3/ITF/hP 1.B, Thrombin, Thrombomodulin, thymidine
phosphorylase (TP), thymosin beta 15, tissue cytosol ferritins,
tissue polypeptide antigen (TPA), tissue polypeptide specific
antigen (TPS), Vascular Endothelial Growth Factor-B (VEGF-B),
Vascular Endothelial Growth Factor-C (VEGF-C), Vascular Endothelial
Growth Factor receptor-1(VEGFR1), Vascular Endothelial Growth
Factor receptor-2 (VEGFR2), and Vascular Endothelial Growth Factor
receptor-3 (VEGFR3).
[0073] Some Genes are overexpressed and can be found by
differential display, including e.g. Claudin-7,
Zinc-alpha-2-glycoprotein, Apolipoprotein B, B94, EST (R08988),
Thrombospondin (THBS1), FGF-1, NGAL/Lipocalin 2, EST (N77731),
BS247 [Abbott Labs WO 9922027], AIB-1. Post translational
modifications can be identified in proteins, including e.g.
Tyrosine phosphorylation, ErbB-2, and EGFR. Absence of key tumor
suppression markers include e.g. mammastatin and maspin.
[0074] Turning now to the figures, FIG. 1 provides a ductal access
device comprising 10 an access tube 12 having a distal end 14, at
least one lumen therethrough, and dimensions which permit
introduction of the distal end through the ductal orifice and
positioning a distal end thereof distal to the ductal sphincter of
a human breast, e.g., typically having an outer access tube
diameter in the range from 0.5 mm to 1 mm, preferably being tapered
within this range over a length from 2 to 3 mm. The device can also
comprise means on the access tool for positioning the distal end
distal to the ductal sphincter. The device can have a stop 16 or
other means to prevent the device from penetrating the duct too
deeply. Alternatively, the tube could have a shoulder or other
enlargement to block penetration at a point at which it is
desirable to stop the penetration of the tool; or, alternatively, a
collar can be placed or built onto the external portion of the
access tube to prevent penetration beyond the collar.
[0075] The means provided to position the device distal to the
ductal sphincter can comprise marks 18 on the access portion of the
device to indicate a penetration depth as indicated in FIG. 2.
Additionally, it may be desired that the device is anchored just
distal to the ductal sphincter once the distal tip has passed
through the ductal sphincter. This may be facilated by any number
of means, including, e.g. placing a small nob or hub 20 on the tube
12 which acts a stop to resist removing the distal tip once the nob
has passed by the ductal sphincter and resides distal to it.
Anchoring the distal tip of the ductal access device distal to the
ductal sphincter may also be accomplished by placing the distal tip
to a depth beyond the ductal sphincter and inflating a balloon (not
shown) to anchor the device below the ductal sphincter during the
infusion and collection procedure.
[0076] The device may also comprise a stop or hub or other means
for keeping the tube accessing the duct from penetrating too far,
and for positioning the access tube distal to the ductal sphincter.
Thus, the device may include a positioning means comprising a stop
element formed or attached to the tube. The stop element has
dimensions which prevent further insertion of the tube into the
duct, and the stop is positioned on the tube so that the distal tip
will be located distal to the ductal sphincter when the device is
fully inserted up to the stop, thus ensuring correct positioning of
the tube in the duct relative to the ductal sphincter. The access
lumen will terminate in at least one port for fluid infusion and/or
collection, and the port is preferably placed at the end of the
distal tip of the device so that it opens in a distal (axial)
direction relative to the access tube 12, and the port is
preferably located relative to the stop element so that the port
resides distal to the ductal sphincter when the stop element
engages the nipple. The stop element can comprise a hub attached to
a proximal end of the tube, wherein the hub has a width which is
greater than the diameter of the tube so that a shoulder is formed
at a junction between the tube and the hub.
[0077] The access device can also be anchored to the external
portions of the accessed breast by any means capable of
accomplishing the anchoring. During the procedure it is important
that the access device not slip out of the duct. Portions of the
device that are external to the access breast duct can be affixed,
strapped, tethered, taped, or otherwise anchored to the breast
during the procedure in order to ensure that the device does not
slip out of the duct. Such anchoring also provides the practitioner
with better control of the device parts if part or all of the
device is anchored, and therefore does not need to be held by the
practitioner or an assistant.
[0078] In a preferred aspect of the catheter design, the access
tube 12 will branch into an infusion arm 22 and collection arm 24.
The infusion arm 22 terminates in a connector 26 which removably
connects to a syringe 28 or other pressurized source of wash fluid.
The collection arm 24 will preferably include a valve 30 and an end
connector 32 for removable attachment to a collection apparatus,
such as a vial, tube, tray, microliter plate, another syringe, or
the like. As discussed below, the collection arm will usually be
closed, e.g., with valve 30, during infusion of the wash fluid.
Preferably, both the infusion arm lumen and collection arm lumen
will be connected to a single lumen within the access tube 12.
[0079] Turning now to FIG. 3, a preferred embodiment of the device
is shown in a single lumen ductal access device 30 having a tube 32
that accesses the duct and through which fluid is infused, and from
which fluid is collected or drawn up out of the duct. A hub 34 is
connected to an infusion tube 36 from which fluid is infused into
the access tube 32 and a collection tube 38 from which fluid is
collected from the access tube. The collection tube 38 is
preferably attached to the hub 34 at a position no closer to the
access tube 32 than the infusion tube 36. Preferably, the
collection tube 38 is located further away from the access tube 32
than is the infusion tube 36. A stylet 40 is optionally provided to
facilitate introduction of the access tube through a ductal orifice
into a ductal lumen. The stylet 40 will pass through a pneumostatic
seal at a proximal end 42 of the hub 34 so that the stylet can be
removed after positioning of the access tube 32 and prior to the
infusion/collection of the wash fluid.
[0080] Fluid is infused into the hub 34 and into the duct until
resistance is met during the infusion. At this time, it is assumed
that the duct is filled. The infusion lumen can be closed and the
fluid allowed to remain in the duct for a preselected time. During
this preselected time, the breast may be massaged and squeezed to
stimulate mixing of the wash fluid and ductal fluid, and also
ultimately to encourage the fluid to leave the duct and enter the
manifold hub. The collection lumen is opened and the breast
squeezed to urge the fluid to progress through the access tube in
the hub. If desired, when cloudy return fluid is seen in the hub
(which is preferably transparent or includes a transparent window),
the infusion lumen can be opened and fluid infused to push the
fluid that has collected in the hub into the collection lumen and a
waiting collection receptacle. Alternatively, and possibly
additionally, aspiration pressure can be applied at the collection
lumen to aspirate any fluid remaining in the hub into the
collection receptacle. The process is repeated either following
another infusion of fluid into the duct or by another round of
squeezing to encourage return and collection of the infused
fluid.
[0081] The stylet 40 can be made of metal or hard plastic and may
have a tapered and/or an atraumatic tip for gently probing and
accessing a breast duct. Preferably, a tapered tip 44 will extend
distally of the access tube 32 as the tube is introduced. After
access of the duct is complete, the stylet 40 can be withdrawn and
the access tube positioned so that its distal end is distal to the
ductal sphincter. The dilator receiving portion at the proximal end
of the device can be a water tight membrane or sheath to provide a
sterile environment in the hub even with penetration and withdrawal
of the dilator, and to provide an appropriate amount of resistance
so that the probe can be manipulated into the out of the duct and
the access tube. The dilator stated in FIG. 3 is removably received
in the access tube and has a distal tip which is positionable
through the access tube to extend from the distal end of the access
tube. In addition to providing tapered access, the stylet 40
selectively stiffens the access tube to further ease introduction
into and through the ductal orifice. The access tube 32 may have an
outer diameter in a range from about 0.25 mm to 1.25 mm with an
inner lumen diameter in the range from 0.2 mm to 1.2 mm.
[0082] As illustrated in FIG. 3A, the hub 34 can have an infusion
connector 46 providing a fluid outlet path into the lumen of the
tube 32, and a collection connector 48 providing a fluid outlet
path from the lumen of the tube. These infusion and collection
connectors are preferably isolated from each other so that the
fluid may be infused through the infusion connector and
simultaneously removed through the collection connector. The
distance l.sub.1 between the infusion port 46 and access tube 32 is
preferably minimized, usually being 1 cm or less, while the
distance l.sub.2 between the infusion port 46 and collection port
48 may be from 0 mm to 2 cm, preferably being 1 cm or less. While
illustrated on opposite sides of the hub 34, the infusion port 46
and collection port 48 may have any relative radial orientation,
with an alignment of both ports on the same side of the hub being
presently preferred.
[0083] FIG. 4A depicts a single lumen access tube 50 accessing the
breast duct D and positioned with its distal end 14 distal to the
ductal sphincter S of the breast duct. FIG. 4B depicts filling the
breast duct D and allowing the fluid to remain in the duct for a
preselected time. FIG. 4C depicts removing the infused fluid mixed
with ductal fluid through the access tube that remains in the duct
during the filling of the duct and collecting of the fluid. FIG. 4D
depicts a catheter 52 having infusion and collection arms 54 and 56
exterior to the accessed duct for separately infusing fluid into
the duct and collecting fluid from the duct. The invention provides
a device having an access tube, a distal end, a single lumen,
dimensions to permit insertion of the device distal to the ductal
sphincter. The device also has an infusion connector, a collection
connector, and that the infusion and collection connectors are
isolated.
[0084] FIG. 6 depicts an alternative ductal access device 72 with
stop cocks 74 and 76 to control the fluid flow into and out from an
accessed duct. A syringe 78 for infusing fluid into the duct is
connected to one arm 80 of an access tube 82. A collection tube 86
is connected to another arm 84 connected to the access tube 82. The
access tube may have single, dual, or multiple lumens as described
elsewhere herein. FIG. 7 depicts the device of FIG. 6 accessing a
breast duct BD. Fluid may be infused from syringe 78 with stopcock
76 open and stopcock 74 closed. After infusing a desired volume as
set forth above, and optionally massaging the breast, stopcock 74
may be opened and fluid collected in passive receptacle 86 (FIG. 6)
or actively withdrawn using a second syringe 88 to apply a vacuum
through the collection arm 84.
[0085] FIGS. 8 and 8A-8C illustrate a dual lumen catheter 100
having lumens 102 and 104 that access the breast duct. A reduced
diameter region 106 accesses the breast duct and the catheter
resides in the duct at a depth of about 3.5 cm. The reduced
diameter portion 106 of the catheter has three lumens 103, 105, and
a third central lumen which receives a fixed stiffening wire 108
(FIG. 8B). The proximal part of the catheter 100 that does not
access the breast duct is depicted in cross section in FIG. 8A
having interior lumen 104 and an annular lumen 102 both which
provide fluid flow (either collection or infusion) during use of
the catheter to retrieve cellular material from the breast duct.
Proximal connector 110 is near a proximal end of the proximal
portion and branches into an infusion arm 112 and aspiration arm
114, each connected to one of the lumens 102 and 109. The distal
portion 106 and proximal portion meet at shoulder 116 where the
lumens 102 and 104 make a transition to annular lumens 103 and 105
(FIG. 8B). Distal tip 120 may be atraumatic for entry into a ductal
orifice and ductal lumen. Usually, side ports 122 will be formed on
the distal section 106 to permit fluid inflow and outflow from the
lumens 103 and 105, respectively.
[0086] FIGS. 9A and 9B depict two formats of transition between a
distal end of an access tube 130 which accesses the breast duct and
the proximal end which resides outside the breast duct. Shoulder
132 in FIG. 9A has a graduated transition, and shoulder 134 in FIG.
9B has a stepped transition.
[0087] The invention also provides systems and kits 60 for
collecting cellular material from a breast duct. FIG. 5 depicts a
system comprising an access device 62, such as a single lumen
catheter having infusion and collection arms 64 and 66 outside the
ductal access portion of the catheter, optionally a premeasured
solution to infuse into the duct and instructions for use of the
catheter and wash fluid to access a breast duct and retrieve
cellular material, e.g., contained in syringe 70. The system 60
includes instructions setting forth any of the methods described
herein, such as the method for obtaining cellular material from a
human breast milk duct comprising introducing a ductal access
device having at least one lumen therethrough into a duct,
introducing a wash fluid through the access device lumen into the
milk duct, wherein a volume of at least 2 ml is present within the
duct for a preselected time; and collecting at least a portion of
the wash fluid from the duct through the lumen of the access
device. Wash fluid can be included in the kit and can comprise for
example, saline, phosphate buffered saline, a nonabsorbable fluid,
an isotonic solution, an osmotic solution, a hypotonic solution, a
hypertonic solution, a protein, a colloid, a sugar, a polymer,
mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, polyethyleneglycol (PEG),
maltodextrin, dextran (e.g. dextran 70), hydroxyethyl starch, fluid
gelatin, a synthetic colloid, an antibody, a binding protein, or
albumin.
[0088] Other ductal access systems available comprise a ductal
access device as a container holding a premeasured volume of ductal
wash fluid. The ductal access device comprises an access tube
having a distal end, at least one lumen therethrough, and
dimensions which permit introduction of the distal end through a
ductal orifice and positioning a distal end thereof distal to the
ductal sphincter of a human breast. The device may also comprise a
means on the access tube for positioning the distal end distal to
the ductal sphincter. The container can comprise a syringe for
connection to the first side port. The pre-measured volume is in
the range from 2 ml to 100 ml. The wash fluid can comprise for
example, saline, phosphate buffered saline, a nonabsorbable fluid,
an isotonic solution, an osmotic solution, a hypotonic solution, a
hypertonic solution.a protein, a colloid, a sugar, a polymer,
mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, polyethyleneglycol (PEG),
maltodextrin, dextran (e.g. dextran 70), hydroxyethyl starch, fluid
gelatin, a synthetic colloid, an antibody, a binding protein, or
albumin.
[0089] Another ductal access system comprises a ductal access
device comprising an access tube having a distal end, a single
lumen therethrough, and dimensions which permit introduction of the
distal end through a ductal orifice and positioning a distal end
thereof distal to the ductal sphincter, an infusion connector
providing a fluid flow path into the lumen of the access tube; and
a collection connector providing a fluid outlet path from the lumen
of the access tube, said infusion and collection connectors being
isolated from each other so that fluid may be infused through the
infusion connector and simultaneously removed through the
collection connector, the system also including a container holding
a premeasured volume of ductal wash fluid. The container can
comprise a syringe for connection to the first side port, and the
premeasured volume can be in the range from 2 ml to 100 ml. The
fluid can comprise saline, phosphate buffered saline, a
nonabsorbable fluid, an isotonic solution, an osmotic solution, a
hypotonic solution, a hypertonic solution, a protein, a colloid, a
sugar, a polymer, mannitol, sorbitol, glucose, glycerol, sucrose,
raffinose, fructose, lactulose, sodium chloride, polyethyleneglycol
(PEG), maltodextrin, dextran (e.g. dextran 70), hydroxyethyl
starch, fluid gelatin, a synthetic colloid, an antibody, a binding
protein, or albumin.
[0090] Another ductal access system can comprise a ductal access
device comprising a hub having an internal elongate manifold, a
lower port at a bottom of the manifold, and first and second side
ports spaced above the lower port; and an access tube having a
distal end, a proximal end, a lumen therethrough, and dimensions
which permit introduction of the distal end through a ductal
orifice and a positioning a distal end thereof distal to the ductal
sphincter of the human breast, wherein the proximal end of the tube
is attached to the lower port of the hub, the ductal access system
comprising also a container holding a premeasured volume of ductal
wash fluid. The container can comprise a syringe for connection to
the first side port. The pre-measured volume is in the range from 2
ml to 100 ml. The ductal access fluid can comprise, e.g., saline,
phosphate buffered saline, a nonabsorbable fluid, an isotonic
solution, an osmotic solution, a hypotonic solution, a hypertonic
solution, a protein, a colloid, a sugar, a polymer, mannitol,
sorbitol, glucose, glycerol, sucrose, raffinose, fructose,
lactulose, sodium chloride, polyethyleneglycol (PEG), maltodextrin,
dextran (e.g. dextran 70), hydroxyethyl starch, fluid gelatin, a
synthetic colloid, an antibody, a binding protein, or albumin.
[0091] Additionally, a ductal access system is provided comprising
a ductal access catheter comprising a catheter body having a distal
end and a proximal end and including at least a distal portion and
a proximal portion, wherein the distal portion has a
cross-sectional geometry which can be inserted through a ductal
orifice into a ductal lumen of a human breast, wherein the proximal
portion has a cross-sectional geometry which inhibits insertion
through the ductal orifice and into the ductal lumen; and wherein
the catheter body has at least an infusion lumen and an collection
lumen each of which has a distal port near a distal end of the
distal portion and a proximal connector near a proximal end of the
proximal portion, the ductal access system comprising also
instructions for use setting forth a method for lavage of a ductal
network in a human breast including introducing a wash fluid
through the infusion lumen into the ductal network and withdrawing
the wash fluid and substances borne by the wash fluid from the
ductal network through the collection lumen.
[0092] At least one of the lumens of the breast duct access device
can have a means to control the fluid flow through that lumen. The
inflow lumen can be connected to a syringe or other infusion
mechanism for infusing lavage fluid into the breast duct. The
outflow lumen can be connected to a collection tube, a collection
syringe, or other collection means for collecting the lavage fluid
after it has mixed with the ductal fluid in the breast duct. The
means to control the fluid flow in a lumen can be, e.g. a stopcock,
valve or other control unit that is capable of closing or opening a
port of the lumen. The lumens themselves may be compressible or
pinchable with fingers or clamps or other pinching or compressing
mechanism. A stopcock may be attached at a lumen of a dual lumen
catheter to control the fluid flow through that lumen. An inflow
lumen may have an inflow stop cock to control fluid flow through an
inflow port. An outflow lumen may have an outflow stop cock to
control fluid flow through an outflow port. The device can have
stopcocks (or other means to control fluid flow) on both an inflow
and an outflow lumen. These control units, valves or stopcocks are
capable of operating separately, e.g. so that when an inflow port
is opened, an outflow port can be closed, etc. Thus, patterns of
control of the fluid flow in a lavage procedure of a breast duct
can include, e.g. an open inflow when an outflow is closed, an open
inflow when an outflow is opened, a closed inflow when an outflow
is opened, and a closed inflow when an outflow is closed. The
catheter may have the graduated duct probe attached to it at the
distal end for accessing the duct. Where the probe is unattached to
the catheter, and is used for dilating the orifice, the catheter
can have a tip appropriate for accessing the dilated duct upon
removal of the probe.
[0093] Lavage fluid can be a saline solution, e.g. normal saline,
or phosphate buffered saline (PBS), or other fluid capable and
suitable for washing a body duct. The lavage fluid will generally
be biocompatible and nontoxic to the patient. The lavage fluid can
further comprise additives, e.g. gas, particles or other fluids.
These additives to the lavage fluid may have various purposes,
however, during a lavage procedure, the preeminent purpose will
generally be to increase a recovery of fluid and/or cellular
material, and/or molecular species from the ducts. Thus, such gas
may provide a cleansing action on the ductal walls for example,
encouraging ductal epithelial cells located e.g. in a lesion in the
duct to shed and be retrievable during the lavage procedure.
Similarly, particle additives may serve to encourage fluids,
cellular material and/or molecular species to follow the particles
in the flow of lavage fluid through the ducts and be retrieved in
the lavage procedure. Such additives as detergents, e.g. agents
tending to form micelles for collecting ductal contents including
cells and molecular species may provide additional yields of cells,
molecular species and fluids in a lavage procedure. The gas can be
ambient air or a related product, and the lavage fluid can comprise
the air mixed in with the fluid for delivery into the duct. The
presence of air or other gas may serve to increase the retrieval of
cells and fluid as compared to a procedure conducted using lavage
fluid alone. The air can be bubbled into the fluid, or introduced
into the fluid mixture by other standard means. The air may also be
mixed into the lavage fluid as the lavage fluid is delivered into
the duct, e.g. where the infusion port allows for delivery of both
air and lavage fluid into the inflow lumens where the two mix and
both are delivered to the accessed ducts.
[0094] The lavage fluid can further contain other agents that may
aid in the retrieval of fluid or cells or both from the duct or may
serve some other useful purpose in the procedure. For example, the
lavage fluid may include or be preceded by or followed by such
other agents that may aid in the retrieval of fluid or cells or
both from the duct, or may serve some other useful purpose in the
procedure. Such other agents can be, for example, an oncotic and/or
osmotic agent capable of increasing the amount of collectable fluid
in the ductal lumen, or a detergent that can help wash out more
cells, or an agent that may help detach more cells from the duct
wall into the ductal lumen (e.g. trypsin, collagenase, or EDTA).
The agent can be an oncotic agent and/or an osmotic agent or both.
Oncotic and osmotic agents are agents that retain fluid around them
or draw fluid to them. The agent can be soluble, e.g. soluble in a
suitable solvent, including e.g. water, buffered water, or a saline
solution. Preferably the solvent is biologically compatible with
mammals. Suitable solvents will be those that both effectively
dissolve the agent and are not toxic to a mammal. The agent can be
a molecular species including e.g. a protein, colloid, sugar, or
polymer. The agent can be mannitol, sorbitol, glucose, glycerol,
sucrose, raffinose, fructose, lactulose, sodium chloride, albumin,
polyethyleneglycol (PEG), maltodextrin, dextran (e.g. dextran 70),
hydroxyethyl starch, fluid gelatin, or a synthetic colloid. Agents
including e.g. mannitol, sorbitol, PEG, glycerol are described in
THE MERCK INDEX, 12.sup.th ed. 1996, Whitehouse Station, N.J.
Others, including maltodextrin, dextran and others are available
from Aldrich Chemical Co. in Milwaukee, Wis. or Sigma Chemical Co.
in St. Louis, Mo. The molecular weight of a suitable oncotic agent
can be determined as optimally within the range of the molecular
weights of suitable oncotic agents available.
[0095] Where the agent in the lavage fluid is a protein, the
protein can be a binding protein or an antibody. The binding
protein can be albumin. The antibody can be capable of binding an
epitope found in a breast duct, e.g. an epithelial cell surface
marker or cancer cell marker, etc. Where the agent is a protein,
the protein is of a molecular weight in the neighborhood of albumin
or higher, so that it is capable of acting as an oncotic agent in
the lumen of the milk duct. Suitable antibodies are commercially
available. Also the agent can be a mixture of osmotic and/or an
oncotic agents. The oncotic agent and/or osmotic agent can comprise
a mixture of any two or more osmotic and/or oncotic agents, e.g.
mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, albumin, polyethyleneglycol
(PEG), maltodextrin, dextran (e.g. dextran 70), hydroxyethyl
starch, fluid gelatin, an antibody or a synthetic colloid.
Preferably the agent is not toxic to a mammal, particularly not
toxic to a human. The agent can be an agent not capable of freely
diffusing into or beyond the cells that line the milk ducts of the
breast. The agent can also be an agent not capable of absorption
into the cells within the duct. For example, the agent can have a
molecular weight large enough to make absorption or diffusion into
the breast duct lining, cells or interstitial space beyond the
lining improbable.
[0096] The method provides that the catheter is used to access the
breast duct after being primed (i.e. filled) with lavage fluid with
both outflow and inflow ports closed. The outflow stop cock can be
opened and the fluid allowed to infuse into the duct from the
outflow to flush the outflow port at the catheter tip and make it
ready to receive the ductal fluid and wash fluid into the outflow
lumen during the lavage procedure. The outflow port at the stopcock
is then closed by closing the outflow stopcock. The inflow port is
opened by opening the inflow stopcock. Wash fluid is infused into
the breast duct until resistance is met. The amount of this first
infusion bolus will vary depending on the size of the breast duct
being infused. The inflow port is then closed by closing the inflow
stopcock. The breast is massaged by applying manual external
pressure on the breast tissue. The outflow lumen is opened and the
breast is massage and squeezed and fluid is collected in the
collection receptacle attached to the outflow lumen. The process
can be repeated several times. Subsequent to the first larger bolus
of wash fluid, lesser amounts of wash fluid can be infused into the
duct and collected in the outflow collection receptacle as just
described.
[0097] A practitioner desirous of increasing a yield of fluid and
cells from a lavage of a patient's breast ducts, and/or desirous of
retrieving fluid and cells from distal regions of the ductal
architecture, can massage the breast of the patient once the fluid
has been infused into the duct. The fluid is infused into the duct
(with the outflow port closed) to a point of resistance and then
the inflow port is closed. The breast can be massaged at this point
to effect a mixing of the ductal fluid with the lavage fluid, and
to generally provide some gentle disruption of ductal cells in the
duct and allow them to enter the fluid mix. The outflow port can be
opened at this point, and allow the massaging can continue, it can
be supplemented with a squeezing or compressing of the breast, i.e.
from the base of the breast upwards towards the nipple in order to
encourage as much fluid to escape via the outflow lumen and into
the collection receptacle.
[0098] Modifications to the method of lavage can include that the
patient is seated during the lavage procedure, rather than the
standard or classic supine (face up) position. In addition, the
patient may be lavaged in a prone position, face down, with nipples
and breast down. The prone face down position takes advantage of
gravity and allows the breast ducts to drain into the collection
receptacle during the procedure when the outflow port is open.
Thus, the lavaging procedure can include infusing the breast duct
with a wash fluid through an open inflow lumen while an outflow
lumen is closed; closing the inflow lumen when the duct is filled;
squeezing or massaging the breast or both; and opening the outflow
lumen to collect the wash fluid.
[0099] The cells collected can comprise ductal epithelial cells;
the ductal fluid collected can comprise molecular and cellular
material. Analysis of the ductal epithelial cells and/or the
molecular and cellular material in the ductal fluid can proceed as
described below discussing the diagnostic methods possible of these
collected materials. The collected cells and fluid and fluid
components can be analyzed, e.g. as described or suggested herein.
The lavage fluid including the ductal cells can be analyzed for
diagnostic purposes. Conditions in a breast milk duct that are
desirable to diagnose include a cancer or precancer condition. The
precancer condition can include atypical ductal hyperplasia (ADH)
or low grade ductal carcinoma in situ (LG-DCIS). The diagnostic
agent may also have the ability to diagnose other breast related
conditions, including, e.g. fibrotic, cystic or conditions relating
to lactation. Diagnostic agents can be mixed with the ductal fluid
(either in the lavage procedure, or after the fluid is
collected).
[0100] The diagnostic agents can include tags for detecting lesions
or other abnormalities or characteristic anatomical or molecular
identities in the breast ducts, including e.g. chemical tags or
antibodies. The tags may provide the capacity for visualizing the
location of a lesion, including, e.g. fluorescent tags, or
biotinylated tags. Antibodies can also be tagged so that the
binding antibody is identifiable. Antibodies can be whole
antibodies, or parts of antibodies including, e.g. Fab fragments,
heavy and/or light chain fragments, single chain antibodies and
other modified antibodies commonly known about and used in the
field of antibody-assisted diagnosis. Diagnostic antibodies or
other tags can be to a number of markers, including e.g. the
following cancer markers that are exemplary and may be used to
analyze the breast duct condition. Standard assay procedures for
identifying the markers can be used. analyzed for the presence of
soluble factors or other components that might indicate the
presence of cancerous or precancerous ductal epithelial cells in
the duct. The epithelial cells retrieved from the breast duct can
be analyzed for protein markers, nucleic acid markers, chromosomal
abnormalities, or other characteristic changes that would signal
the presence of cancerous or precancerous cells. In addition, other
cells found in the duct can also be analyzed, e.g. for an increase
or decrease in these cells as compared to normal ductal fluid, or
for qualities of these cells themselves. Thus, the condition of the
breast duct can be analyzed e.g. for soluble protein content or
presence of other ductal fluid components, including also secreted
products of ductal epithelial cells) or the ductal epithelial cells
themselves can be analyzed, for example, for cell morphology, for
protein markers, for nucleic acid markers, and for biochemical
markers.
[0101] In addition, any of the cells of the duct can be analyzed
for morphological abnormalities in cell components, including, e.g.
morphological abnormalities of the nucleus, cytoplasm, Golgi
apparatus or other parts of a cell. The cells can be analyzed for
whether they do or don't aggregate (e.g. in clumps) or by making
comparisons of the ductal epithelial cells with other cell types
retrieved in the ductal fluid (e.g. macrophages, lymphocytes, foam
cells and other possible components of ductal fluid). The ductal
epithelial cells can be analyzed for their molecular contents or
the morphology of the ductal epithelial cells, including, e.g.
protein markers, nucleic acid markers, biochemical markers in the
cells or on the cell surfaces or for any evidence of neoplasia.
[0102] In addition to some markers discussed and/or articles or
books cited on breast cancer and breast precancer markers,
including markers listed in Porter-Jordan and Lippman, "Overview of
the biological markers of breast cancer", Hematology/Oncology
Clinics of North America vol. 8 (1):73-100, 1994), the following
cancer markers are listed here as exemplary and may be used as well
as other markers to analyze the condition of a breast duct,
including analysis of the ductal contents (including fluid and
cells). Standard assay procedures for identifying the markers can
be used, including antibodies or other binding partners, labels,
stains, pattern analysis (for cells and cell components), and in
general any other chemical or visual identification techniques.
[0103] Markers that are presently being studied by researchers
presently include, carcinoma embryonic antigen (CEA), prostate
specific antigen (PSA) Erb B2 antigen, gross cystic disease fluid
protein-15 (GCDFP-15), and lactose dehydrogenase (LDH). For CEA see
Imayama et al, Cancer 1996, 78(6):1229-34; Inaji et al, Cancer
1987,60(12):3008-13; Mori Int Conger Seer 1989, 807:211-8; Inaji,
et al, An To Kagaku Ryoho 1991, 18(2):313-7; Yayoi, et al Gan To
Kagaku Ryoho 1994, 21 Suppl 2:133-9; Mori, et al Jpn J Clin Oncol
1989,19(4):373-9; Foretova, et al Proc Annu Meet Am Soc Clin Oncol
1995,14:A101; and Nishiguchi, et al Rinsho Byori 1992,40(1):67-72.
For PSA see Foretova, Garber Lancet 1996,347(9015):1631; Sauter et
al, Cancer Epidemiology, Biomarkers & Prevention. 5(12):967-70,
1996; Sauter and Daly (1996) Proc Annu Meet Am Assoc Cancer Res
37:A1458; and Foretova and Garber (1996) Proc Annu Meet Am Assoc
Cancer Res 37:A1446. For Erb B2 see Motomura (1995) Breast Cancer
Res and Treat 33:89-92; and Inaji et al (1993) Tumour Biol
14:271-8. For GCDFP-15 see Petrakis et al (1994) Proc Annu Meet Am
Assoc Cancer Res 35:A1698. For LDH see Mannello et al (1995) Cancer
76:152-4; and Kawamoto (1994) Cancer 73:1836-41.
[0104] Chromosomal abnormalities in ductal epithelial cells can
also provide information and act as a marker to identify cancer or
precancer as described in Mark et al (1999) Cancer Genet Cytogenet
108:26-31; Lundlin and Mertens (1998) Breast Cancer Res Treat
51:1-15; Newsham (1998) Am J Pathol 153:5-9; Larson et al (1998) Am
J Pathol 152:1591-8; Adelaide et al (1998) Genes Chromosomes Cancer
22:186-99; Fejzo et al (1998) Gene Chromosome Cancer 22:105-113;
Dietrich et al (1998) Hum Pathol 12: 1379-82; Cavalli et al (1997)
Hereditas 126:261-8; Adeyinka et al (1997) Cancer Genet Cytogenet
97:119-21; Afify and Mark (1997) Cancer Genet Cytogenet 97:101-5;
Brenner and Aldaz (1997) Prog Clin Biol Res 396: 63-82; Mark et al
(1997) Ann Clin Lab Sci 27:47-56; and Fabian et al 1993 J. Cellular
Biochemistry 17G:153-16.
[0105] In addition, exemplary markers are described in Masood,
(Prediction of recurrence for advanced breast cancer. Traditional
and contemporary pathologic and molecular markers) Surgical
Oncology Clinics of North America. 4(4):601-32, 1995;
Lopez-Guerrero et al (1999) J Hematother 8(1):53-61; Marjumdar and
Diamandis (1999) Br J Cancer 79(9-10):1594-602; Balleine et al
(1999) Br J Cancer 79 (9-10):1564-71; Houston et al (1999) Br J
Cancer 79(7-8):1220-6; Nikolic-Vukosavljevic et al (1998) Tumori
84(6):691-4; Maguire et al (1998) Int J Biol Markers 13(3):139-44;
Steams et al (1998) Breast Cancer Res Treat 52(1-3):239-59;
Eiriksdottir et al (1998) Eur J Cancer 34(13):2076-81, and U.S.
Pat. No. 5,169,774. Many known breast cancer markers are discussed
and described in readily available medical textbooks on breast
cancer. Other markers are also listed herein.
[0106] The morphology of the cells or cellular contents retrieved
in the ductal fluid and wash fluid may also be examined. The
cellular contents can include, e.g. protein, nucleic acid, or other
molecular markers in the cells. Cell morphology can serve to
establish whether the ductal epithelial cells are normal (i.e. not
precancerous or cancerous or having another noncancerous
abnormality), precancerous (i.e. comprising hyperplasia, atypical
ductal hyperplasia (ADH) or low grade ductal carcinoma in situ
(LG-DCIS)) or cancerous (i.e. comprising high grade ductal
carcinoma in situ (HG-DCIS), or invasive carcinoma). Analysis of
cell contents may serve to establish similar staging as established
by morphology, capturing generally a progression of a precancerous
or cancerous condition in the cells.
[0107] Administering fluid to the ductal lumen for the purpose of
collecting that fluid mixed with the fluid from the duct is
complicated by the fact that absorbable wash fluids are partly
absorbed into the breast from the duct. Thus, the fluid retrieved
is less than that infused, even considering that it includes the
ductal fluid that was residing in the duct. Administering an agent
in the wash fluid that is capable of increasing or maintaining the
fluid volume in the duct is a great advantage to the process. Thus,
the invention provides administering a nonabsorbable fluid or a
fluid that actually draws fluid to it, e.g. an oncotic or osmotic
fluid in the process of collecting fluid from the duct.
Administering the nonabsorbable fluid has the advantage also of
providing the practitioner with a way to monitor or standardize the
ductal fluid and cellular return in any given volume of fluid
infused and retrieved. For example 10 ml of the nonabsorbable fluid
is administered to the duct, and 9.5 ml of that fluid is collected.
Maybe 100 epithelial clusters are contained in the fluid collected.
This information can be noted, and during future procedures on that
same duct can be compared. The advantage of using a nonabsorbable
is that the ductal fluid yield may be increased with the retrieval
of most or all of the infused fluid, and the practioner will be
able to keep track of the amount infused versus the amount
collected.
[0108] A nonabsorbable fluid can be used in order to provide a
standardization to the process so that the amount infused can be
correlated with the amount collected, knowing that since the fluid
cannot be absorbed in the duct, and collecting of all or most of
the fluid that is infused is possible.
[0109] Identification of the location of the ducts prior to
accessing them can be made as described in PCT application to the
Regents of the University of California at Los Angeles filed Sep.
15, 1998 to Barsky et al entitled "Methods and Kits for Identifying
Ductal Orifices in a Nipple", or U.S. Ser. No. 09/153,564 filed
Sep. 15, 1998 to Barsky et al.
[0110] The agent is an agent capable of in effect increasing the
amount of collectable fluid in the ductal lumen. Thus the agent can
be a nonabsorbable agent or fluid or an oncotic agent and/or an
osmotic agent or a combination of two or all three. Oncotic and
osmotic agents are agents that retain fluid around them or draw
fluid to them. The agent can be soluble, e.g. soluble in a suitable
solvent, including e.g. water, buffered water, or a saline
solution. Preferably the solvent is biologically compatible with
mammals. Suitable solvents will be those that both effectively
dissolve the agent and are not toxic to a mammal.
[0111] The agent can be a molecule including e.g. a protein,
colloid, sugar, or polymer. The agent can be mannitol, sorbitol,
glucose, glycerol, sucrose, raffinose, fructose, lactulose, sodium
chloride, albumin, polyethyleneglycol (PEG), maltodextrin, dextran
(e.g. dextran 70), hydroxyethyl starch, fluid gelatin, or a
synthetic colloid. Agents including e.g. mannitol, sorbitol, PEG,
glycerol are described in THE MERCK INDEX, 12.sup.th ed. 1996,
Whitehouse Station, N.J. Others, including maltodextrin, dextran
and others are available from Aldrich Chemical Co. in Milwaukee,
Wis. or Sigina Chemical Co. in St. Louis, Mo. The molecular weight
of a suitable oncotic agent can be determined as optimally within
the range of the molecular weights of suitable oncotic agents
available.
[0112] Where the agent is a protein, the protein can be a binding
protein or an antibody. The binding protein can be albumin. The
antibody can be capable of binding an epitope found in a breast
duct, e.g. an epithelial cell surface marker or cancer cell marker,
etc. Where the agent is a protein, the protein is of a molecular
weight in the neighborhood of albumin or higher, so that it is
capable of acting as an oncotic agent in the lumen of the milk
duct. Suitable antibodies are commercially available.
[0113] Also the agent can be a mixture of osmotic and/or an oncotic
agents. The oncotic agent and/or osmotic agent can comprise a
mixture of any two or more osmotic and/or oncotic agents, e.g.
mannitol, sorbitol, glucose, glycerol, sucrose, raffinose,
fructose, lactulose, sodium chloride, albumin, polyethyleneglycol
(PEG), maltodextrin, dextran (e.g. dextran 70), hydroxyethyl
starch, fluid gelatin, an antibody or a synthetic colloid.
[0114] The agent can be an agent not capable of freely diffusing
into or beyond the cells that line the milk ducts of the breast.
The agent can also be an agent not capable of absorption into the
cells within the duct. For example, the agent can have a molecular
weight large enough to make absorption or diffusion into the breast
duct lining, cells or interstitial space beyond the lining
improbable.
[0115] Whether an agent is capable capable of increasing or at
least maintaining the amount of collectable fluid (with relation to
the amount of fluid infused) in the ductal lumen can be determined
by experimentation to identify whether collectable fluid in the
duct is increased upon administration of an agent as compared to
administration of a control isotonic solution to a neighboring
control duct. Likewise the best volume and concentration of the
agent can be determined by a comparison of the amount of
collectable fluid yielded with a change in a variable such as a
volume or concentration of agent administered. The agents including
nonabsorbable fluid and/or oncotic and/or osmotic agents to be
tested can be delivered to the duct of a human, rat, rabbit, pig or
other appropriate mammal, and the ductal fluid can be collected.
Where the fluid yield is greater than control fluid collected from
a neighboring duct (after injection of a control solution,
preferably of equal volume as the tested solution), that agent is
suitable for use in the method. The increased fluid amount should
be at least 50% and more preferably close to 100% of an increase of
fluid collectable from the ducts that are compared. In the case
where the practitioner seeks to increase the amount of fluid
collected from the amount infused, the fluid yield from the duct
administered with the agent being tested can be several fold that
of the control fluid yield. Where the goal is merely to provide for
a collection fluid amount that is close to the amount infused, the
parameters for success are that the amount of fluid collected from
the duct after infusion of a set aliquot of fluid is closer to the
amount infused that would have been possible if the infusion fluid
had been an absorbable fluid such as saline. Such a comparison can
be tested by doing a control infusion and collection in a duct
using e.g. saline and then repeating the procedure in the same duct
using a nonabsorbable fluid, e.g. a PEG containing fluid or the
like.
[0116] The appropriate concentration and volume of oncotic agent
and/or osmotic agent in solution injected into a duct can be
determined by routine experimentation including cannulation or
catheterization of mammalian nipples (e.g. rat, rabbit, pig or
human nipples) to determine at which concentration and volume the
agent in solution yields the most volume of fluid collectable from
the duct as compared to the fluid collectable from a control duct.
Experiments can be designed for testing a variety of oncotic and/or
osmotic agents, concentrations, volumes, and mixtures of agents in
all varieties of mammals having breast ducts.
[0117] Fluid collected from the milk ducts, can include
constituents of biological fluids, e.g. those typically found in
breast duct fluid, e.g. water, cells, cellular markers, molecular
markers, nucleic acids, proteins, cellular debris, salts, or
organic molecules. These constituents can be analyzed by any
appropriate method depending on the practitioner's purposes in
obtaining the fluid.
[0118] The fluid can comprise cells including e.g. epithelial cells
and abnormal cells. The cells can be analyzed for cellular,
protein, nucleic acid, or other molecular markers or for shape or
other abnormalities. Analysis of the cells can provide diagnostic
or prognostic information for an evaluation of the condition of the
breast or breast ducts. Removal of cells can be conducted in the
presence of the agent, and preferably the action of the osmotic
and/or oncotic agent provides for removing cells that can be
analyzed.
[0119] The invention includes a kit for increasing the amount of
fluid collectable from a milk duct of a breast comprising an
nonabsorbable agent and/or an osmotic agent and/or an oncotic
agent, a medical tool for delivering the agent to the ductal lumen,
and instructions for use. The nonabsorbable agent and/or oncotic
and/or osmotic agent can be those described herein or other
comprising like properties and/or functions in a breast duct. The
medical tool can be any tool that enables delivery of such agent.
The instructions can direct a protocol for administration including
how to administer the agent, how much time to wait before
collecting the fluid, how to collect the fluid, and how to analyze
the fluid collected.
[0120] The retrieved fluid can comprise constituents of the breast
milk duct fluid, e.g. including water, cells, cellular markers,
molecular markers, nucleic acids, proteins, cellular debris, salts,
or organic molecules. Analyses can be made that identify molecular
or cellular markers, cellular characteristics, e.g. by cytology,
and for making any other assessment of any of the constituents of
the fluid. Cells that are retrieved and analyzed can be epithelial
cells or abnormal cells.
[0121] Multiple lumen ductal access catheters, having more than one
lumen in the access portion of the catheter, according to the
present invention will comprise a catheter body having a distal end
and a proximal end and including at least a distal portion and a
proximal portion. The catheter will have at least two continuous
lumens extending through both the proximal and distal portions. The
lumens can be fluid carrying, and fluid can pass from the proximal
portion to the distal portion of one lumen and from the distal
portion to the proximal portion of a second lumen. The distal
portion has a cross-sectional geometry which can be inserted
through a ductal orifice into a ductal lumen of a human breast for
the purpose of lavaging the breast duct. The proximal portion has a
cross-sectional geometry which inhibits insertion through the
ductal orifice and into the ductal lumen thereby placing limits on
the extent that the catheter penetrates the breast duct during the
lavage procedure. The lumens of the catheter are an infusion lumen
and an aspiration or collection lumen. Each lumen has a distal port
near the distal end of the distal portion of the catheter. One
distal port is for infusing liquid into the duct (a port on the
infusion lumen). The other distal port is for aspirating or
collecting fluid from the duct (a port on the aspiration lumen).
The catheters also have a proximal connector near a proximal end of
the proximal portion for connecting e.g. to a fluid receptacle that
holds fluid for infusion into the duct (e.g. a wash or lavage
fluid) and for connecting to a collection receptacle for collecting
the contents of the duct that are aspirated in the aspiration lumen
(e.g. a syringe that can both aspirate and collect the ductal fluid
and/or ductal contents).
[0122] The ductal access catheter will have a total length in the
range of from about 2-cm to about 60-cm, usually about 30 cm to
about 45-cm. The length of the proximal portion will typically be
in the range of about 15-cm to about 50 cm, more typically in the
range from about 30 cm to about 40 cm. The distal portion will
typically be in the range from about 2.5-cm to about 8 cm, more
typically in the range from about 3.0 cm to about 5.5 cm.
[0123] The proximal and distal portions will preferably be joined
to each other with an intermediate zone between them to accommodate
the difference in cross-sectional geometry between the proximal and
distal portions. The intermediate zone can be a stepped decrease in
cross-sectional geometry from proximal to distal portions, or may
be a gradual decrease in cross-sectional geometry from proximal to
distal portions. The body segments can be joined in any
conventional manner to each other (and/or to the either end of the
element creating the intermediate zone) including methods such as
heat fusion, adhesive bonding, coextrusion, or the like. In the
exemplary embodiment, the distal and proximal portions will be
coextruded and the coextrusion process will generate the
intermediate zone in accommodating the differential cross-sectional
geometry of the proximal to the distal portions.
[0124] The catheter can have the distal portion of the catheter
body stiffened over at least a part of its length to facilitate
insertion through the ductal orifice and into the ductal lumen. The
stiffening effect can be created by insertion of a third lumen in
the distal portion, the third lumen comprising a wire. The wire can
be made of some relatively stiff metal, e.g. tungsten or steel. The
stiffened distal portion of the catheter body can have an average
bending stiffness in the range from about 0.010 inch-lbs to about
0.5 inch-lbs. Typically the bending stiffness of the distal portion
will be about 0.105 inch-lbs.
[0125] The catheter may be composed of any biologically compatible
polymeric resins or metal having suitable characteristics when
formed into the tubular catheter portions. Exemplary materials
include polyvinyl chloride, polyethers, polyamides, polyethylenes,
polycarbonate, polyurethanes, copolymers thereof and the like. The
distal portion may be formed of the same or different material as
the proximal portion. Although a stiffening wire may be placed in
the distal portion, if the stiffening wire is not present, the
distal portion may be composed of materials that are slightly more
stiff than the materials that compose the proximal portion.
Optionally, the distal body portion may be reinforced with a metal
or polymeric braid or other conventional reinforcing layering.
[0126] The distal portion will be sufficiently rigid to permit
axial positioning of the distal tip in a ductal orifice with the
distal portion extending either partly or wholly into the breast
ductal lumen. The distal portion will typically have a hardness in
a durometer range at least greater than that of the proximal
portion, and thus generally greater than 75 D. The hardness of the
distal portion thus may be a range from about 70 D to about 90 D.
The proximal portion will be more flexible and less stiff and also
less hard than the distal portion. The durometer of the proximal
portion outer tubing can be in a range from about 45 A to about 100
A, and typically about 80 A. The inner tubing of the proximal
portion can have a durometer in the range from about 50 D to about
75 D, and typically about 63 D. The flexibility of the proximal
portion provides the catheter with the advantages that the distal
portion (which is stiffer) can be inserted into the breast duct,
meanwhile the proximal portion can connect at its hubs with
infusion or collection apparatus and not kink during the placement
of the distal portion in the breast duct. Additionally, the
flexibility of the proximal portion provides the advantage that
once the distal portion is placed in the breast duct the catheter
will have less tendency to pull out of the duct. The stiffness of
the distal portion benefits the procedure by allowing access into
the orifice of the duct and the duct itself, an action that
requires a probe-like quality of the distal portion and distal tip
in order the duct to be accessed successfully.
[0127] The catheter body may further comprise other components,
such as radiopaque fillers; colorants; reinforcing materials;
reinforcement layers, such as braids and helical reinforcement
elements; or the like. In particular it would be possible to
reinforce the distal portion in order to enhance its duct
penetration or probe-like capabilities while optionally limiting
its wall thickness and outside diameter so that the catheter can
easily access even ducts with small ductal orifices.
[0128] The cross-sectional geometry of the distal portion of the
catheter body will be smaller than the cross-sectional geometry of
the proximal portion. The cross-sectional geometry of the distal
portion provides that the distal portion can be inserted into a
breast duct orifice and through the orifice into the breast duct
lumen. The distal portion of the catheter body has a maximum width
in the range from 0.008 inches to 0.050 inches. The distal portion
of the catheter body has a generally tubular structure with a
diameter in the range from about 0.008 inches to about 0.035
inches. The proximal portion has a cross-sectional geometry which
inhibits insertion of the proximal portion into the ductal orifice
and the ductal lumen. Thus, the proximal portion of the catheter
body has a minimum width in the range from about 0.023 inches to
about 0.028 inches. The proximal portion of the catheter body has a
generally tubular structure with a diameter in the range from about
0.030 inches to about 0.10 inches. The proximal diameter is greater
than the distal diameter by at least about 0.010 inches.
[0129] The region between the proximal and distal portions of the
catheter body provides for the reduced diameter moving from the
proximal to the distal portions. The transition preserves the fluid
flow capability and communication in the lumens between the
proximal and distal portions and can provide a place to anchor or
lodge a wire or stiffening lumen that extends longitudinally in the
distal portion. The transition may be stepped, abrupt, or somewhat
gradual, provided it allows the proximal portion to retain its
function of inhibiting insertion of the catheter into the duct
beyond the length of the distal portion.
[0130] The ductal access catheter body can comprise at least an
infusion lumen and an aspiration lumen each of which has a distal
port near a distal end of the distal portion. At least one of the
distal aspiration port and the distal infusion portion can be
disposed on a side of the distal portion of the catheter body. Thus
one port can be a side port and one port can be and end port. The
distal aspiration port and the distal infusion port can both be
located on the side of the distal portion of the catheter body.
Thus, both ports for both lumens can be side ports. The distal
aspiration port and the distal infusion port can be axially
aligned. Thus, the side ports can be located e.g. opposite each
other on the on the distal portion of the catheter body at the
longitudinal position. For example, both side ports can be located
about 2.5 cm from the distal tip. The distal aspiration port and
the distal infusion port can be axially spaced apart. Thus, the
side ports can be located at different longitudinal positions on
the distal portion, for example one port can be located about 2.0
cm from the distal tip and one port can be located about 2.5 cm
from the distal tip. The side ports themselves may be round or oval
or any other geometric shape conducive to fluid flow either into
the duct or out from the duct. The diameter of the ports can be
that diameter which is suitable to achieve a desired flow rate into
the duct or aspiration or collection rate out from the duct. Thus,
the diameters of the ports can be in a range from about 0.015
inches (0.038 mm) to about 0.022 inches (0.056 mm), most typically
in a range from about 0.016 inches (0.041 mm) to about 0.020 inches
(0.051 mm). One side port can be larger or smaller than the other,
especially where such differential port size provides a desired
flow rate into or out from one of the lumens, or an overall lavage
efficiency of infusion and aspiration or collection of lavage and
ductal fluid.
[0131] The catheter body can include an atraumatic distal tip. The
tip can be contoured and/or rounded to reduce or eliminate trauma
to the duct upon entry through the ductal orifice and penetration
into the ductal lumen. The tip may also be fashioned to reduce or
eliminate trauma upon withdrawal of the tool from the duct after
the lavage procedure is completed. The tip can be composed of a
soft polymeric material, e.g. including polyvinyl chloride,
polyethers, polyamides, polyethylenes, polyurethanes, copolymers
thereof and the like. The tip can have a diameter in the range from
about 0.012 inches (0.031 mm) to about 0.020 inches (0.051 mm),
more typically a diameter in the range from about 0.014 inches
(0.036 mm) to about 0.018 inches (0.046 mm). The length of the tip
(extending from the distal end of the distal portion of the
catheter) can be in a range from about 0.25 cm to about 2.5 cm,
more typically in the range from about 0.50 cm to about 1.8 cm. The
invention also provides a method for lavage of a ductal network in
a human breast comprising providing a catheter as any described
above for performing the lavage procedure. The distal portion of
the catheter is inserted through a ductal orifice and into a distal
lumen of the ductal network. A wash fluid is introduced through the
infusion lumen into the ductal network. The wash fluid can be, e.g.
saline or phosphate buffered saline, or any biocompatable fluid
suitable for washing a breast duct lumen. The wash fluid and
substances borne by the wash fluid are withdrawn from the ductal
network through the aspiration lumen. The various features of the
catheters described above can serve to facilitate the practice of
the lavage procedure. For example, the narrow distal tip provides
the catheter the ability to penetrate the ductal orifice and move
the catheter into the ductal lumen for performing the lavage
procedure; the larger diameter of the proximal portion inhibits the
catheter from passing too deeply into the duct, and stops the
penetration of the catheter at the place where the distal portion
ends and the proximal portion begins; the atraumatic tip provides
the catheter the ability to penetrate the duct without trauma to
the tissue walls of the ductal lumen; the stiffening material
placed in at least a part of the distal portion of the catheter
(e.g. a stiffening wire or a supporting braid or the like) provides
the practitioner with stiffness to better control the entry and
further penetration of the catheter into the ductal lumen; the
ports on the lumens provide the catheter the ability to infuse
liquid into the duct from the infusion lumen and the ability to
aspirate or collect fluid from the duct into the aspiration lumen;
and where the ports are side ports, the presence of side ports may
better facilitate the function of the ports for infusing and
collecting to and from the duct.
[0132] The invention further provides a ductal access system
comprising any of the catheters describe herein and instructions
for use setting forth a method for lavage of a ductal network in a
human breast including introducing a wash fluid through the
infusion lumen into the ductal network and withdrawing the wash
fluid and substances borne by the wash fluid from the ductal
network through the aspiration lumen, e.g. as described for the
method above.
EXAMPLES
[0133] 1. Collecting Cells and Cellular Material Using Single Lumen
Ductal Access Device
[0134] Device as depicted in FIG. 3 was used to access breast ducts
of patients A, B, C, D, E, F, G, and H. Before ductal access
patient's nipple was cleaned with alcohol, and dekeratinized with
cerumetix. An aspiration cup was placed on the nipple and areola
and the patient's nipple was aspirated to identify the breast duct
and to collect fluid for a comparison with the fluid retrieved from
inside the duct. A small quantity of fluid was observed on the
nipple surface after aspiration and this fluid was collected with
one or more capillary tubes placed in contact with the fluid. The
aspiration fluid was preserved in a preservative solution for cells
for analysis later.
[0135] Ducts that yield fluid were accessed using a dilator that
extended from the device depicted in FIG. 3, and once the duct was
accessed by the access tube, the dilator was withdrawn. The
collection tube was closed, and the system including the infusion
tube and manifold were primed with fluid. A total of from 10 ml to
about 25 ml of saline infusion fluid was infused into the duct
until resistance was felt in the infusion syringe. The assumption
made at that point was that the duct was filled with the infusion
fluid. The infusion tube was closed and the collection tube opened.
The breast was massaged and then squeezed and cloudy fluid was
caused to enter the hub and begin to exit the collection tube. To
encourage the fluid to exit, the infusion tube was opened and
additional infusion fluid was pushed into the hub, causing more
cloudy fluid to exit the collection tube. The following fluid
amounts refer to the procedure with Patient A, When a volume of
about 11.5 ml of fluid was collected, the collection tube was
closed and more fluid infused until a resistance was felt in the
duct. More fluid was infused to refill the duct. The collection
tube was opened, and infusion tube was closed and the breast was
massaged and squeezed to encourage more fluid to enter the hub and
exit the collection tube. Additionally, the fluid was encouraged to
leave the hub with an injection of fluid from the infusion lumen.
About 6 ml was collected from the second filling.
[0136] The results of the nipple aspiration (NAF), first filling
and second filling are reported below for patient A in the Table I.
Patient's B, C, and D also have NAF results compared to results
using the single lumen catheter, as depicted in Table I. Patients
E, F, G and H have yields solely with respect to access and
retrieval using the single lumen catheter. Epithelial cell clusters
are defined as clusters of cells having greater than 10 epithelial
cells per cluster.
1TABLE I Total volume Epithelial Clusters Sample Collection
collected (>10 cells/cluster) Patient A; NAF (nipple aspiration
>0.1 ml 1 epithelial cluster duct R2 fluid) lavage with single
lumen 17.5 ml 23 epithelial clusters Patient B; NAF -- 0 epithelial
clusters duct L6 lavage with single lumen -- 31 epithelial clusters
Patient C; NAF 0.2 ml 0 epithelial clusters duct R1 lavage with
single lumen 6 ml 27 epithelial clusters Patient D; NAF <0.1 ml
0 epithelial clusters duct on lavage with single lumen 7 ml 101
epithelial clusters left nipple Patient E; lavage with single lumen
11 ml 3 epithelial clusters duct L6 Patient F; lavage with single
lumen 10.5 ml 12 epithelial clusters duct L6 duct L7 lavage with
single lumen 7 ml 7 epithelial clusters duct R1 lavage with single
lumen 21 ml 6 epithelial clusters duct R2 lavage with single lumen
7 ml 5 epithelial clusters Patient G; lavage with single lumen 6 ml
400 epithelial clusters duct L6 duct R1 lavage with single lumen
8.5 ml 350 epithelial clusters Patient H; lavage with single lumen
11 ml 154 epithelial clusters duct L6 duct R1 lavage with single
lumen 7 ml 131 epithelial clusters
[0137] 2. Comparative Study: Pig Pelt Lavage with StopCock Catheter
vs. Catheter without StopCocks
[0138] In order to test the efficiency of the stop cock catheter, a
comparison was run comparing the amount of fluid and cells
retrieved from a catheter which did not have stop cocks (or on
which both stop cocks were kept open during the entire procedure),
and a catheter having a stop cock on the inflow lumen and a stop
cock on the outflow lumen that are opened and closed alternately
during the procedure (in the manner specified below). Two
experiments (experiment I and experiment II) were conducted using
modified procedures A and B to test modifications to the basic
principles of the comparison. The combined results show the
increased efficiency of using the stop cock catheter (in the manner
described below in the procedure A portion of experiments I &
II) to retrieve a larger volume of infused wash fluid and a larger
number of ductal cells from the lavaged breast milk duct.
[0139] A dual lumen catheter having a support wire is used for the
tests. The catheter also has a stopcock on the inflow lumen and a
stopcock on the outflow lumen. Frozen pig pelts were purchased from
Yosemite Meats, located in Menlo Park, Calif. Procedure A was
conducted using the stopcocks controlling the opening and closing
of the inflow and outflow lumens at ports located in the stopcocks.
Procedure B was conducted with both the inflow and outflow lumens
open during the entire procedure. Experiments I and II were
conducted as follows:
[0140] Procedure A included the following steps:
[0141] 1. Catheter was inserted into a duct. Both inflow and
outflow lumens were primed with wash fluid, and the ports closed
(by placing the respective stopcocks in a closed position). The
catheter was placed in the duct.
[0142] 2. The outflow port was opened, and 1 ml of phosphate
buffered saline (PBS) was infused into the duct to flush out the
outflow lumen. The outflow port was closed.
[0143] 3. The inflow port was opened and PBS was infused into the
duct until resistance to infusion was met. The inflow port was
closed.
[0144] 4. The outflow port was opened. The breast was massaged and
squeezed. The outflow fluid was collected.
[0145] 5. The outflow port remained opened, the inflow port was
opened, and 0.5 ml of fluid was infused into the inflow to flush
out the outflow lumen. The inflow port was closed and the fluid
collected in the collection syringe attached to the outflow
lumen.
[0146] 6. The outflow port was closed and about 1 ml of PBS was
infused. The inflow port was closed. The outflow port was opened.
The breast was massaged and squeezed, and the fluid collected.
Steps 5 and 6 were repeated until about 3 ml of fluid was
collected.
[0147] Procedure B included the following steps:
[0148] 1. A dual lumen catheter (with both inflow and outflow ports
open) was inserted into the duct.
[0149] 2. The duct was infused with PBS until the fluid flow into
the duct met resistance.
[0150] 3. The duct was lavaged using massaging and squeezing
technique as the fluid was collected in the collection receptacle
(located at the end of the outflow lumen).
[0151] 4. More PBS was infused (each time about 1 ml) and the
massaging, squeezing and collecting proceeded. The procedure was
repeated until about 3 ml was collected.
[0152] Procedure A (one-way flow procedure) included the following
steps:
[0153] 1. Both the inflow and outflow lumens were primed with PBS.
The inflow and outflow ports were closed and the catheter inserted
into the pig duct to a depth of about 1.5 cm.
[0154] 2. The outflow port was opened and 1 ml of PBS was infused
into the duct to flush out the outflow lumen.
[0155] 3. The outflow port was closed. The inflow port was opened.
Fluid was infused in the inflow port (about 4-5 ml) until
resistance was felt. The inflow port was closed.
[0156] 4. The breast was massaged with both ports closed.
[0157] 5. The outflow port was opened and the breast squeezed to
collect fluid in the collection receptacle until no more fluid
comes out.
[0158] 6. The inflow port was opened and about 0.5 ml of fluid was
infused to flush out the outflow lumen.
[0159] 7. The outflow port was closed and about 1 ml of PBS was
infused into the duct in the inflow lumen.
[0160] 8. The inflow port was closed. The outflow port was opened,
and the breast massaged and squeezed to collect the outflow fluid.
Steps 6, 7, and 8 were repeated until the collection volume totaled
3 ml.
[0161] Procedure B included the following steps:
[0162] 1. Both the inflow and outflow lumens of a dual lumen
catheter were primed with PBS. The catheter tip was inserted into a
breast duct of a pig pelt to a depth of about 1.5 cm. About 1 ml of
PBS was infused into the duct through the outflow lumen to flush
out the outflow lumen.
[0163] 2. PBS was infused into the inflow port until resistance was
felt.
[0164] 3. The breast was massaged and squeezed as the duct was
lavaged with PBS. Fluid flowing to the outflow lumen was
collected.
[0165] 4. More PBS was infused into the duct (each time about 1 ml)
and the duct ravaged, (using massaging and squeezing) and the fluid
collected in the collection receptacle. The procedure was repeated
until about 3 ml of fluid was collected.
[0166] The fluid was used to prepare Cytospin.RTM. slides by taking
10 ul of collected fluid (plus 90 ul of PBS), using a cytospin
machine to place the fluid on a slide. The slides are air-dried and
Diff Quik.RTM. stained. The results are shown below in Table II. In
all cases 3 ml of fluid was collected, but the infusion volume
varied as shown in the table.
2TABLE II Procedure A Procedure B infusion infusion nipple vol cell
density nipple vol cell density experiment I X2 15 ml 55% Z2 13 ml
45% X3 18 ml 65% X1 10 ml 35% Z3 13 ml 40% Z1 13 ml 45% Z5 12 ml
55% experiment II X1 14 ml 60% Z1 7 ml 50% X2 12 ml 65% Z2 6 ml 50%
X3 13 ml 65% Z3 6 ml 40% X4 11 ml 55% Z4 6 ml 35% average 13.5 ml
58% 8.7 ml 43%
[0167] The results indicated a 35% increased cell yield using
procedure A over procedure B.
[0168] 3. Optimal Stop Cock Catheter Usage for Retrieving Cells
[0169] A procedure was developed that appeared to optimize the
potential yield of cells from the ductal fluid retrieved was a
catheter-based lavage procedure of a breast duct. Using pig pelts
the following technique resulted in maximized cell yield from a set
volume of collected fluid.
[0170] 1. Inflow and outflow lumens of a dual lumen catheter having
stop cocks on both lumens were primed with PBS. The inflow and
outflow ports were closed and the catheter inserted into the duct
to a depth of about 1.5 cm.
[0171] 2. The outflow port was opened and 1 ml of PBS was infused
into the duct from the outflow lumen to flush out the outflow
lumen.
[0172] 3. The outflow port was closed (using the stopcock) and the
inflow port was opened. PBS was infused into the duct until
resistance was felt (about 4-5 ml of PBS). The inflow port was
closed.
[0173] 4. The breast was massaged while both the inflow and outflow
ports were closed (using the stopcock controls).
[0174] 5. The outflow port was opened and the breast was massaged
and squeezed, and the outflow fluid was collected until no more
fluid came out.
[0175] 6. The outflow port was closed and the inflow opened, and
more fluid (about 1 ml) was allowed to infuse into the duct.
[0176] 7. The inflow port was closed and the outflow port was
opened. The breast was massaged and squeezed to collect the
fluid.
[0177] 8. Steps 6 and 7 were repeated until a total of 3 ml of
fluid was collected.
[0178] This procedure was found to generate the best cell density
collected in the 3 ml of fluid, and also eliminates one step from
previous procedure A.
[0179] 4. Mannitol Solution Introduced into Breast Ducts of Live
Rabbit Results In Increased Ductal Fluid Collection
[0180] The objective of these experiments was to test the effects
of the introduction of a solution containing mannitol on the
secretion of fluid from the breast ducts of live rabbits. New
Zealand rabbit #3242, female, from Kraelik Farm in CA weighing 4.1
kg was used. The rabbit was anesthetized by injection of 200 mg of
ketamine and 40 mg of Zylazine. A second injection of 100 mg of
ketamine and 20 mg of xylazine was made 2 hours later to maintain
the rabbit in a deep plane of anesthesia. The thorax and abdomen of
the rabbit was shaved to expose the breasts and nipples.
[0181] A single lumen blue color catheter (O.D. 0.23" ID 0.017;
O.D. at the tip 0.011"-0.012") was inserted into a duct in each
nipple. Three nipples were tested, and 2 ducts per nipple were
accessed with a catheter. The nipples were identified A, B, and
C.
[0182] A duct on nipple A was injected with 0.20 ml of a 12.5%
solution of D-Mannitol in H.sub.2O (available from Sigma Chemicals,
St. Louis, Mo. cat# M-9546 lot 6710402: C.sub.6H.sub.14O.sub.6 FW
182.2) with a single catheter. The control duct on nipple A was
injected with 0.20 ml of phosphate buffered saline (PBS). A
microfuge tube was attached to the end of each catheter to collect
out flow liquid. Ten minutes later 0.2 ml of a 12.5% solution of
D-Mannitol in H.sub.2O was injected into the first duct and the
second duct was injected with 0.20 ml of phosphate buffered saline,
for a total volume injected in each duct of 0.40 ml.
[0183] A duct on nipple B was injected with 0.5 ml of a 12.5%
solution of D-Mannitol in H.sub.2O with a single catheter. The
control duct on nipple B was injected with 0.50 ml of phosphate
buffered saline (PBS). A microfuge tube was attached to the end of
each catheter to collect out-flow liquid.
[0184] A duct on nipple C was injected with 0.7 ml of a 12.5%
solution of D-Mannitol in H.sub.2O with a single catheter. The
control duct on nipple C was injected with 0.70 ml of phosphate
buffered saline (PBS). A microfuge tube was attached to the end of
each catheter to collect out flow liquid.
[0185] About an hour after the fluid containing mannitol or PBS was
injected into the ducts via the catheters, the microfuge tubes were
checked for whether any fluid was returned. The results are
summarized in the following Table III:
3TABLE III nipple duct solution recovery notes A A1 0.4 ml mannitol
310 ul liquid fluid was a milky color 12.5% A A2 0.4 ml PBS none --
B B1 0.5 ml mannitol 490 ul liquid fluid was a milky color 12.5% B
B2 0.5 ml PBS 240 ul liquid fluid was a milky color C C1 0.7 ml
mannitol 280 ul liquid fluid was a milky color 12.5% C C2 0.7 ml
PBS none --
[0186] Davidson green dye (1 ul) was added to each microfuge tube
containing fluid for the purpose of taking a picture. The rabbit
was euthanized by IV injection of supersaturated KCl. PBS (1.5 ul)
was added to each collection. The cells were spun onto Shandon
coated slide using megafunnel and cytospin-3 machine (Shandon, Inc.
located in Pittsburgh, Pa.) at a speed of 1500/per minute for 15
minutes. The cells were fixed on the slide in 95% ethanol for 10
minutes. The cells were stained using Hematoxylin and Eosin (HE)
method of cytology of collected fluid. The results of the cellular
analysis are in Table IV:
4TABLE IV Nipple A/ Nipple B/ Nipple B/ Nipple C/ duct A1 duct B1
duct B2 duct C1 A few ductal A few ductal A few ductal A few ductal
cell clusters cell clusters cell clusters cell clusters and
scattered and scattered and scattered and scattered histocytes and
histocytes and histocytes and histocytes and apocrine apocrine
apocrine apocrine metaplastic metaplastic metaplastic metaplastic
cells cells cells cells
[0187] The observations made from this experiment are that fluid
can be collected from three out of three ducts injected with
mannitol solution; that fluid could be collected from 1 out of 3
ducts injected with PBS solution, and with approximately 50% less
volume in the ducts where fluid was collected. There were cells
detected from the fluid collected from each duct. The cell
morphology looked similar between the mannitol and the PBS injected
ducts.
[0188] All publications and patent applications cited in this
specification are herein incorporated by reference as if each
individual publication or patent application were specifically and
individually indicated to be incorporated by reference. Although
the foregoing invention has been described in some detail by way of
illustration and example for purposes of clarity of understanding,
it will be readily apparent to those of ordinary skill in the art
in light of the teachings of this invention that certain changes
and modifications may be made thereto without departing from the
spirit or scope of the appended claims.
* * * * *