US4994387A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process and medium for cloning and long-term serial cultivation of human endothelial cells
Assignee: Thomas Jefferson University
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Filing: 1988-06-17
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Granted: 1991-02-19
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Expires: 2008-06-17
Abstract
Abstract Vigorous growth of endothelial cells from human umbilical vein endothelial cells in vitro is achieved using a culture medium containing endothelial cell growth factor and heparin and/or dextran sulfate. Cell population doubling times of 13 to 21 hours for 60 to 85 populations doublings are obtained, and cloned human endothelial cell strains are established having similar proliferative capacities.
Inventors
- 👤 Elliot M. Levine
- 👤 Sandor S. Shapiro
- 👤 Bruce E. Jarrell
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 10:15
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/069; C12N2501/00; C12N2501/90; C12N2501/91; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/948
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
84 words
A culture medium for enhancing proliferation and increasing the lifespan of human endothelial cell cultures which comprises a cell growth medium, and effective amounts of endothelial cell growth factor (ECGF) and a material selected from the group consisting of heparin, dextran sulfate, and mixtures thereof to increase the population doublings of human umbilical vein endothelial cells (HUVE cells) to at least about 42 prior to senescence and to reduce the doubling time of said HUVE cells so as not to exceed about 21 hours.
Claim 2
Dependent Claim
84 words
A culture medium for enhancing proliferation and increasing the lifespan of human endothelial cell cultures which comprises a cell growth medium, and effective amounts of endothelial cell growth factor (ECGF) and a material selected from the group consisting of heparin, dextran sulfate, and mixtures thereof to increase the population doublings of human umbilical vein endothelial cells (HUVE cells) to at least about 42 prior to senescence and to reduce the doubling time of said HUVE cells so as not to exceed about 21 hours.
Claim 3
Dependent Claim
45 words
A culture medium according to claim 1 comprising from about 5% to about 30% fetal bovine serum and from about 70% to about 95% of a cell growth medium, said percentages being by volume based on the total volume of said endothelial cell culture medium.
Claim 4
Dependent Claim
28 words
A cell culture medium according to claim 2 containing from about 10 to about 200 μg/ml of ECGF and from about 9 to about 900 μg/ml of heparin.
Claim 5
Dependent Claim
26 words
A cell culture medium according to claim 3 comprising about 20% fetal bovine serum and containing about 20 μg/ml of ECGF and about 90 μg/ml heparin.
Claim 6
Dependent Claim
29 words
A cell culture medium according to claim 2 containing from about 10 to about 200 μg/ml of ECGF and from about 5 to about 500 μg/ml of dextran sulfate.
Claim 7
Dependent Claim
22 words
A cell culture medium according to claim 5 comprising about 20% fetal calf serum, and containing about 100 μg/ml of dextran sulfate.
Claim 8
Dependent Claim
98 words
A process for enhancing proliferation and increasing the life span of human umbilical endothelial cell cultures which comprises culturing human endothelial cells derived from human umbilical veins in the presence of a cell culture medium containing effective amounts of endothelial cell growth factor (ECGF) and a material selected from the group consisting of heparin, dextran sulfate, and mixtures thereof to increase the population doublings of human umbilical vein endothelial cells (HUVE cells) to at least about 42 prior to senescence and to reduce the doubling time of said HUVE cells so as not to exceed about 21 hours.
Claim 9
Dependent Claim
98 words
A process for enhancing proliferation and increasing the life span of human umbilical endothelial cell cultures which comprises culturing human endothelial cells derived from human umbilical veins in the presence of a cell culture medium containing effective amounts of endothelial cell growth factor (ECGF) and a material selected from the group consisting of heparin, dextran sulfate, and mixtures thereof to increase the population doublings of human umbilical vein endothelial cells (HUVE cells) to at least about 42 prior to senescence and to reduce the doubling time of said HUVE cells so as not to exceed about 21 hours.
References Cited (Backward Citations)
US4443546A
Cited
US4585654A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-02-18 | 1994-08-20 | 1995-02-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-02-18 | 1998-08-20 | 1999-02-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-02-18 | 2002-08-20 | 2003-02-19 | $7700.00 | Window Closed / Lapsed |