US4927762A
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Cell culture medium with antioxidant
Assignee: Cell Enterprises Inc
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Filing: 1988-11-30
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Granted: 1990-05-22
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Expires: 2008-11-30
Abstract
Abstract Cells grown in chemically defined (serum free) media are protected against endogenous oxidizing agents by a nontoxic antioxidant with free thiol groups, at cell densities less than about 10 5 cells/ml, especially DL-penicillamine, N-acetylcysteine, mercaptoproprionic acid, 2-mercaptoethanesulfonic acid, and thiolactate. These agensts are useful in the maintenance and growth of hybridomas in such media.
Inventors
- 👤 Frederick J. Darfler
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 09:49
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/10; C12N5/12; C12N5/16; C12N5/163; C12N2500/00; C12N2500/05; C12N2500/10; C12N2500/20; C12N2500/24; C12N2500/25; C12N2500/30; C12N2500/44; C12N2500/90
Claims Analysis
Total Claims: 31
Claim 1
Independent Claim
51 words
A chemically defined, serum free, albumin-free medium for maintenance or growth of immortal or immortalized cells which comprises a cell-growth promoting amount of a compound having one free thiol group, and which is capable of supporting the growth of immortal or immortalized cells at seed densities less than 10 5 cells/ml.
Claim 2
Dependent Claim
51 words
A chemically defined, serum free, albumin-free medium for maintenance or growth of immortal or immortalized cells which comprises a cell-growth promoting amount of a compound having one free thiol group, and which is capable of supporting the growth of immortal or immortalized cells at seed densities less than 10 5 cells/ml.
Claim 3
Dependent Claim
54 words
The medium of claim 1 wherein the compound has the formula: ##STR2## where R 1 and R 2 are H or CH 3 , R 3 and R 4 are H, NH 2 or NH(CO)CH 3 , and R 5 is COO - , SO 3 - or CH 2 --SO 3 - .
Claim 4
Dependent Claim
35 words
The medium of claim 1 wherein the compound does not have the structure HS--CHZ--CHK--Y where X is H or NH 2 , Y is COO - , and Z is H or CH 3 .
Claim 5
Dependent Claim
12 words
The medium of claim 1 wherein the cell-growth-promoting amount is 0.1-1.0 mmoler/liter.
Claim 6
Dependent Claim
10 words
The medium of claim 1 wherein the compound is thiolactate.
Claim 7
Dependent Claim
35 words
The medium of claim 2 wherein the compound does not have the structure HS--CHZ--CHX--Y where X is H or NH 2 , Y is COO - , and Z is H or CH 3 .
Claim 8
Dependent Claim
49 words
A chemically-defined, serum-free, medium for maintenance or growth of immortal or immortalized cells which comprises a cell growth promoting amount of a compound having one free thiol group, and which is capable of supporting the growth of immortal or immortalized cells at seed densities less than 10 5 cells/ml.
Claim 9
Dependent Claim
49 words
A chemically-defined, serum-free, medium for maintenance or growth of immortal or immortalized cells which comprises a cell growth promoting amount of a compound having one free thiol group, and which is capable of supporting the growth of immortal or immortalized cells at seed densities less than 10 5 cells/ml.
Claim 10
Dependent Claim
53 words
The medium of claim 7 wherein the compound has the formula: ##STR3## where R 1 and R 2 are H or CH 3 , R 3 and R 4 are H, NH 2 or NH(CO)CH 3 and R 5 is COO - , SO 3 - or CH 2 --SO 3 - .
Claim 11
Dependent Claim
35 words
The medium of claim 7 wherein the compound does not have the structure HS--CHZ--CHK--Y where X is H or NH 2 , Y is COO - , and Z is H or CH 3 .
Claim 12
Dependent Claim
12 words
The medium of claim 7 wherein the cell-growth-promoting amount is 0.1-1.0 mmoles/liter.
Claim 13
Dependent Claim
10 words
The medium of claim 7 wherein the compound is thiolactate.
Claim 14
Dependent Claim
35 words
The medium of claim 8 wherein the compound does not have the structure HS--CHZ--CHX--Y where X is H or NH 2 , Y is COO - , and Z is H or CH 3 .
Claim 15
Dependent Claim
42 words
A chemically defined, serum-free medium for the maintenance or growth of immortal or immortalized cells which comprises (a) a cell growth-promoting amount of a compound having one free thiol group, (b) insulin, and (c) transferrin, said medium being otherwise free of proteins.
Claim 16
Dependent Claim
42 words
A chemically defined, serum-free medium for the maintenance or growth of immortal or immortalized cells which comprises (a) a cell growth-promoting amount of a compound having one free thiol group, (b) insulin, and (c) transferrin, said medium being otherwise free of proteins.
Claim 17
Independent Claim
60 words
A method of growing immortal or immortalized cells at seed densities less than 10 5 cells/ml in a chemically defined serum-free medium which comprises inoculating a chemically defined, serum-free medium comprising a cell growth-promoting amount of a compound having one free thiol group with less than 10 5 immortal or immortalized cells/ml medium and growing the cells in that medium.
Claim 18
Independent Claim
60 words
A method of growing immortal or immortalized cells at seed densities less than 10 5 cells/ml in a chemically defined serum-free medium which comprises inoculating a chemically defined, serum-free medium comprising a cell growth-promoting amount of a compound having one free thiol group with less than 10 5 immortal or immortalized cells/ml medium and growing the cells in that medium.
Claim 19
Dependent Claim
12 words
The method of claim 14 in which the medium is also albumin-free.
Claim 20
Dependent Claim
18 words
The method of claim 14 in which insulin and transferrin are the sole protein components of the medium.
Claim 21
Dependent Claim
53 words
The method of claim 14 wherein the compound has the formula: ##STR4## where R 1 and R 2 are H or CH 3 , R 3 and R 4 are H, NH 2 or NH(CO)CH 3 and R 5 is COO - , SO 3 - or CH 2 SO 3 - .
Claim 22
Dependent Claim
35 words
The method of claim 14 wherein the compound does not have the structure HS--CHZ--CHX--Y where X is H or NH 2 , Y is COO - , and Z is H or CH 3 .
Claim 23
Dependent Claim
35 words
The method of claim 17 wherein the compound does not have the structure HS--CHZ--CHX--Y where X is H or NH 2 , Y is COO - , and Z is H or CH 3 .
Claim 24
Dependent Claim
10 words
The method of claim 14 wherein the compound is thiolactate.
Claim 25
Dependent Claim
56 words
A supplemental cell culture medium consisting essentially of transferrin, insulin, ethanolamine, a selenium salt, an unsaturated fatty acid, and a cell-growth-promoting amount of a compound having one free thiol group, where said medium, when added to a basal cell culture medium, supports the growth of hybridoma cells at seed densities of less than 10 5 cells/ml.
Claim 26
Dependent Claim
56 words
A supplemental cell culture medium consisting essentially of transferrin, insulin, ethanolamine, a selenium salt, an unsaturated fatty acid, and a cell-growth-promoting amount of a compound having one free thiol group, where said medium, when added to a basal cell culture medium, supports the growth of hybridoma cells at seed densities of less than 10 5 cells/ml.
Claim 27
Dependent Claim
12 words
The method of claim 14 in which the cells are hybridoma cells.
Claim 28
Dependent Claim
10 words
The method of claim 14 wherein the compound is N-acetylcysteine.
Claim 29
Dependent Claim
12 words
The method of claim 14 wherein the compound is D- or L-penicillamine.
Claim 30
Dependent Claim
10 words
The method of claim 14 wherein the compound is MESNA.
Claim 31
Dependent Claim
10 words
The method of claim 15 wherein the compound is MPA.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-05-21 | 1993-11-20 | 1994-05-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-05-21 | 1997-11-20 | 1998-05-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-05-21 | 2001-11-20 | 2002-05-22 | $7700.00 | Window Closed / Lapsed |