US4757005A
Expired / Lapsed
US
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Method and cell line for obtaining plasminogen activators
Assignee: Bayer Corp
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Filing: 1985-02-22
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Granted: 1988-07-12
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Expires: 2005-02-22
Abstract
Abstract A method of increasing the yield of plasminogen activator from a plasminogen activator-producing cell line by adapting said cell line to grow in a suitable growth medium essentially free of fetal calf serum is disclosed. Also disclosed are cell lines capable of producing relatively large quantities of plasminogen activator, and a chemically defined growth medium suitable for adapting a plasminogen activator-secreting cell line to growth in the absence of fetal calf serum.
Inventors
- 👤 Sham Y. Chan
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 18:53
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6456; C12N1/00; C12R; C12R2001/00; C12R2001/91
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
148 words
A method for increasing the yield of plasminogen activator obtained from plasminogen activator-producing mammalian tumor cells which comprises the steps of: (a) growing said cells in a suitable growth medium supplemented with fetal calf serum, said suitable growth medium composed of a mixture of Waymouth's MB752/1, Dulbecco Minimal Essential Medium and Ham's F-12 medium in a ratio (by weight) of from about 1:1:1 to about 3:1:2, respectively,; (b) passing said cells through a series of said suitable growth media containing decreasing amounts of fetal calf serum, said media supplemented with the growth factors fetuin, bovine serum albumin, insulin, transferrin, 5α-dihydrotestosterone and dexamethasone; (c) growing said cells to confluency in said suitable growth medium essentially free of fetal calf serum and supplemented with said growth factors, said cells producing plasminogen activator at levels greater than that of the initial plasminogen activator-producing cells; and (d) recovering therefrom said plasminogen activator.
Claim 2
Independent Claim
148 words
A method for increasing the yield of plasminogen activator obtained from plasminogen activator-producing mammalian tumor cells which comprises the steps of: (a) growing said cells in a suitable growth medium supplemented with fetal calf serum, said suitable growth medium composed of a mixture of Waymouth's MB752/1, Dulbecco Minimal Essential Medium and Ham's F-12 medium in a ratio (by weight) of from about 1:1:1 to about 3:1:2, respectively,; (b) passing said cells through a series of said suitable growth media containing decreasing amounts of fetal calf serum, said media supplemented with the growth factors fetuin, bovine serum albumin, insulin, transferrin, 5α-dihydrotestosterone and dexamethasone; (c) growing said cells to confluency in said suitable growth medium essentially free of fetal calf serum and supplemented with said growth factors, said cells producing plasminogen activator at levels greater than that of the initial plasminogen activator-producing cells; and (d) recovering therefrom said plasminogen activator.
Claim 3
Dependent Claim
33 words
The method of claim 1 wherein the plasminogen activator-producing cells are mammalian tumor cells selected from the group consisting of melanoma, prostate, breast, colon, ovarian, pancreatic, cervical, rectal, endometrial and fibroblastic tumor cells.
Claim 4
Dependent Claim
31 words
The method of claim 2 wherein the mammalian tumor cells are human carcinoma cell lines selected from the group consisting of melanoma, prostate, breast, colon, ovarian, pancreatic and endometrial cell lines.
Claim 5
Dependent Claim
15 words
The method of claim 3 wherein the tumor cell line is a human prostate adenocarcinoma.
Claim 6
Dependent Claim
30 words
The method of claim 1 wherein the suitable growth medium is an SYC medium composed of a 1:1:1 by weight mixture of Waymouth's MB752/1, Dulbecco MEM and Ham's F-12, respectively.
Claim 7
Dependent Claim
86 words
The method of claim 5 wherein the fetuin is present in a concentration of from 50-150 milligrams per liter; the bovine serum albumin is present in a concentration of from 50-150 milligrams per liter; the insulin is present in a concentration of from 5-10 milligrams per liter; the transferrin is present in a concentration of from 5-40 milligrams per liter; the 5α-dihydrotestosterone is present in concentration of from 5-200 micrograms per liter; and the dexamethasone is present in a concentration of from 50-200 micrograms per liter.
Claim 8
Dependent Claim
81 words
The method of claim 6 wherein the fetuin is present in a concentration of 75 milligrams per liter; the bovine serum albumin is present in a concentration of 75 milligrams per liter; the insulin is present in a concentration of 8 milligrams per liter; the transferrin is present in a concentration of 25 milligrams per liter; the 5α-dihydrotestosterone is present in a concentration of 100 micrograms per liter; and the dexamethasone is present in a concentration of 100 micrograms per liter.
Claim 9
Dependent Claim
24 words
The method of claim 1 wherein the concentration of fetal calf serum is serially decreased by a 2-fold factor through each of said passages.
Claim 10
Independent Claim
155 words
A method for increasing the yield of plasminogen activator obtained from human prostate adenocarcinoma cell line ATCC CRL-1435 which comprises the steps of: (a) growing said cell line in a suitable growth medium supplemented with fetal calf serum, said suitable growth media composed of a mixture of Waymouth's MB752/1, Dulbecco Minimal Essential Medium and Ham's F-12 Medium in a ratio (by weight) of from about 1:1:1 to about 3:1:2, respectively; (b) passing said cell line through a series of said suitable growth media containing decreasing amounts of fetal calf serum, said media supplemented with the growth factors fetuin, bovine serum albumin, insulin, transferrin, 5α-dihydrotestosterone and dexamethasone; (c) growing said cell line to confluency in said suitable growth medium essentially free of fetal calf serum and supplemented with said growth factors, said cell line producing plasminogen activator at levels greater than that of the initial plasminogen activator-producing cell line; and (d) recovering therefrom said plasminogen activator.
Claim 11
Independent Claim
155 words
A method for increasing the yield of plasminogen activator obtained from human prostate adenocarcinoma cell line ATCC CRL-1435 which comprises the steps of: (a) growing said cell line in a suitable growth medium supplemented with fetal calf serum, said suitable growth media composed of a mixture of Waymouth's MB752/1, Dulbecco Minimal Essential Medium and Ham's F-12 Medium in a ratio (by weight) of from about 1:1:1 to about 3:1:2, respectively; (b) passing said cell line through a series of said suitable growth media containing decreasing amounts of fetal calf serum, said media supplemented with the growth factors fetuin, bovine serum albumin, insulin, transferrin, 5α-dihydrotestosterone and dexamethasone; (c) growing said cell line to confluency in said suitable growth medium essentially free of fetal calf serum and supplemented with said growth factors, said cell line producing plasminogen activator at levels greater than that of the initial plasminogen activator-producing cell line; and (d) recovering therefrom said plasminogen activator.
Claim 12
Dependent Claim
30 words
The method of claim 9 wherein said suitable growth medium is an SYC medium composed of a 1:1:1 by weight mixture of Waymouth's MB752/1, Dulbecco MEM and Ham's F-12, respectively.
Claim 13
Dependent Claim
87 words
The method of claim 10 wherein the fetuin is present in a concentration of from 50-150 milligrams per liter; the bovine serum albumin is present in a concentration of from 50-150 milligrams per liter; the insulin is present in a concentration of from 5-10 milligrams per liter; the transferrin is present in a concentration of from 5-40 milligrams per liter; the 5α-dihydrotestosterone is present in a concentration of from 50-200 micrograms per liter; and the dexamethasone is present in a concentration of from 50-200 micrograms per liter.
Claim 14
Dependent Claim
81 words
The method of claim 11 wherein the fetuin is present in a concentration of 75 milligrams per liter; the bovine serum albumin is present in a concentration of 75 milligrams per liter; the insulin is present in a concentration of 8 milligrams per liter; the transferrin is present in a concentration of 25 milligrams per liter; the 5α-dihydrotestosterone is present in a concentration of 100 micrograms per liter; and the dexamethasone is present in a concentration of 100 micrograms per liter.
Claim 15
Dependent Claim
24 words
The method of claim 9 wherein the concentration of fetal calf serum is serially decreased by a 2-fold factor through each of said passages.
Claim 16
Dependent Claim
18 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as PC-3f having ATCC accession number CRL-8539.
Claim 17
Dependent Claim
18 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as PC-3f having ATCC accession number CRL-8539.
Claim 18
Dependent Claim
18 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as PaCa-2f having ATCC accession number CRL-8725.
Claim 19
Dependent Claim
18 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as PaCa-2f having ATCC accession number CRL-8725.
Claim 20
Dependent Claim
19 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as KLE f having ATCC accession number CRL-8726.
Claim 21
Dependent Claim
19 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as KLE f having ATCC accession number CRL-8726.
Claim 22
Dependent Claim
19 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as ML f having ATCC accession number CRL-8724.
Claim 23
Dependent Claim
19 words
A biologically pure culture of a plasminogen activator-secreting cell line designated as ML f having ATCC accession number CRL-8724.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-07-12 | 1992-01-11 | 1992-07-12 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-07-12 | 1996-01-11 | 1996-07-12 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-07-12 | 2000-01-11 | 2000-07-12 | $7700.00 | Window Closed / Lapsed |