Patent US7005252B1 successfully refreshed from Google Patents.
US7005252B1
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US
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Serum free cultivation of primate embryonic stem cells
Assignee: Wisconsin Alumni Research Foundation
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Filing: 2000-03-09
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Granted: 2006-02-28
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Expires: 2020-03-09
Abstract
Abstract Disclosed herein are methods for culturing primate embryonic stem cells. These cells are cultured on a prolonged and stable basis in the presence of exogenously supplied fibroblast growth factor and in the absence of animal serum. Preferably there is also a fibroblast feeder layer. Also disclosed is a culture media containing fibroblast feeder layer and the fibroblast growth factor.
Inventors
- 👤 James A Thomson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 16:30
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0603; C12N5/0606; C12N2500/00; C12N2500/90; C12N2501/00; C12N2501/10; C12N2501/115
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
97 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the primate embryonic stern cells in a culture medium containing albumin, amino acids, vitamins, minerals, at least one transferrin or transferrin substitute, and at least one insulin or insulin substitute, the culture medium being essentially free of mammalian fetal serum and containing exogenously supplied human fibroblast growth factor that is supplied from a source other than just a fibroblast feeder layer, so that the stern cells proliferate in culture and remain undifferentiated in the absence of serum in the medium.
Claim 2
Independent Claim
97 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the primate embryonic stern cells in a culture medium containing albumin, amino acids, vitamins, minerals, at least one transferrin or transferrin substitute, and at least one insulin or insulin substitute, the culture medium being essentially free of mammalian fetal serum and containing exogenously supplied human fibroblast growth factor that is supplied from a source other than just a fibroblast feeder layer, so that the stern cells proliferate in culture and remain undifferentiated in the absence of serum in the medium.
Claim 3
Dependent Claim
16 words
The method of claim 1 , wherein the culture is essentially free of any animal serum.
Claim 4
Dependent Claim
15 words
The method of claim 2 , wherein the culture also comprises a fibroblast feeder layer.
Claim 5
Dependent Claim
16 words
The method of claim 2 , wherein the fibroblast growth factor is basic fibroblast growth factor.
Claim 6
Dependent Claim
25 words
The method of claim 4 , wherein the fibroblast growth factor is human basic fibroblast growth factor which has been produced from a recombinant gene.
Claim 7
Dependent Claim
34 words
The method of claim 5 , wherein the human basic fibroblast growth factor is present in the culture in a concentration of at least 0.1 ng/ml for at least a portion of the method.
Claim 8
Dependent Claim
17 words
The method of claim 2 , wherein the primate embryonic stem cells are human embryonic stem cells.
Claim 9
Independent Claim
122 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the primate embryonic stem cells in a culture medium containing albumin, amino acids, vitamins minerals, at least one transferrin or transferrin substitute, and at least one in or insulin substitute, the culture medium being essentially free of mammalian fetal serum and containing exogenously supplied mammalian fibroblast growth factor that is supplied from a source other than just a fibroblast feeder layer, said culturing step being conducted for over one month with the embryonic stem cells proliferating in culture while maintaining the potential of the stem cells to differentiate into derivatives of endoderm, mesoderm, and ectoderm tissues, and while maintaining the karyotype of the stem cells.
Claim 10
Independent Claim
122 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the primate embryonic stem cells in a culture medium containing albumin, amino acids, vitamins minerals, at least one transferrin or transferrin substitute, and at least one in or insulin substitute, the culture medium being essentially free of mammalian fetal serum and containing exogenously supplied mammalian fibroblast growth factor that is supplied from a source other than just a fibroblast feeder layer, said culturing step being conducted for over one month with the embryonic stem cells proliferating in culture while maintaining the potential of the stem cells to differentiate into derivatives of endoderm, mesoderm, and ectoderm tissues, and while maintaining the karyotype of the stem cells.
Claim 11
Independent Claim
112 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the stem cells in a culture medium containing albumin, amino acids, vitamins, minerals, at least one transferrin or transferrin substitute, and at least one insulin or insulin substitute, the culture medium being essentially free of mammalian fetal serum and in the presence of a fibroblast growth factor capable of activating a fibroblast growth factor signaling receptor, wherein the growth factor is exogenously supplied to the culture from a source other than just a fibroblast feeder layer, so that the stem cells proliferate in culture and remain undifferentiated in the absence of serum in the medium.
Claim 12
Independent Claim
112 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the stem cells in a culture medium containing albumin, amino acids, vitamins, minerals, at least one transferrin or transferrin substitute, and at least one insulin or insulin substitute, the culture medium being essentially free of mammalian fetal serum and in the presence of a fibroblast growth factor capable of activating a fibroblast growth factor signaling receptor, wherein the growth factor is exogenously supplied to the culture from a source other than just a fibroblast feeder layer, so that the stem cells proliferate in culture and remain undifferentiated in the absence of serum in the medium.
Claim 13
Dependent Claim
16 words
The method of claim 9 , wherein the culture is essentially free of any animal serum.
Claim 14
Dependent Claim
15 words
The method of claim 10 , wherein the culture also comprises a fibroblast feeder layer.
Claim 15
Dependent Claim
17 words
The method of claim 10 , wherein the primate embryonic stem cells are human embryonic stem cells.
Claim 16
Independent Claim
139 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the primate embryonic stem cells in a culture medium containing albumin, amino acids, vitamins, minerals, at least one transferrin or transferrin substitute, and at least one insulin or insulin substitute, the culture medium being essentially free of mammalian fetal serum and in the presence of a fibroblast growth factor capable of activating a fibroblast growth factor signaling receptor, wherein the growth factor is exogenously supplied to the culture from a source other than just a fibroblast feeder layer, said culturing step being conducted for over one month with the embryonic stem cells proliferating in culture while maintaining the potential of the stem cells to differentiate into derivatives of endoderm, mesoderm, and ectoderm tissues, and while maintaining the karyotype of the stem cells.
Claim 17
Independent Claim
139 words
A method of culturing primate embryonic stem cells in defined media without serum, the method comprising: culturing the primate embryonic stem cells in a culture medium containing albumin, amino acids, vitamins, minerals, at least one transferrin or transferrin substitute, and at least one insulin or insulin substitute, the culture medium being essentially free of mammalian fetal serum and in the presence of a fibroblast growth factor capable of activating a fibroblast growth factor signaling receptor, wherein the growth factor is exogenously supplied to the culture from a source other than just a fibroblast feeder layer, said culturing step being conducted for over one month with the embryonic stem cells proliferating in culture while maintaining the potential of the stem cells to differentiate into derivatives of endoderm, mesoderm, and ectoderm tissues, and while maintaining the karyotype of the stem cells.
Claim 18
Dependent Claim
81 words
In a method of culturing primate embryonic stem cells without serum, the improvement comprising: culturing the primate embryonic stem cells in a culture free of added mammalian fetal serum but including albumin, vitamins, minerals, insulin, and transferrin, and in the presence of fibroblast growth factor that is exogenously supplied to the culture from a source other than just a fibroblast feeder layer, so that the stem cells proliferate in culture and remain undifferentiated in the absence of serum in the medium.
Claim 19
Dependent Claim
81 words
In a method of culturing primate embryonic stem cells without serum, the improvement comprising: culturing the primate embryonic stem cells in a culture free of added mammalian fetal serum but including albumin, vitamins, minerals, insulin, and transferrin, and in the presence of fibroblast growth factor that is exogenously supplied to the culture from a source other than just a fibroblast feeder layer, so that the stem cells proliferate in culture and remain undifferentiated in the absence of serum in the medium.
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 2009-02-27 | 2009-08-29 | 2010-02-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2013-02-27 | 2013-08-29 | 2014-02-28 | $3760.00 | Window Closed / Lapsed |
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