Culturing mammalian embryonic stem cells in human foreskin fibroblast conditioned medium
First Claim
1. A method of maintaining mammalian pluripotent stem cells in an undifferentiated state comprising culturing the mammalian pluripotent stem cells with a human foreskin fibroblast conditioned medium, wherein said foreskin fibroblast conditioned medium is of non-genetically modified human foreskin fibroblasts, and wherein said foreskin fibroblast conditioned medium comprises at least 4 ng/ml of added basic fibroblast growth factor (bFGF), and being capable of maintaining said mammalian pluripotent stem cells in an undifferentiated, proliferative state through at least 87 passages, thereby maintaining the mammalian pluripotent stem cells in the undifferentiated state.
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Abstract
A cell culture comprising human foreskin cells, the human foreskin cells being capable of maintaining stem cells in an undifferentiated state when co-cultured therewith.
9 Citations
26 Claims
- 1. A method of maintaining mammalian pluripotent stem cells in an undifferentiated state comprising culturing the mammalian pluripotent stem cells with a human foreskin fibroblast conditioned medium, wherein said foreskin fibroblast conditioned medium is of non-genetically modified human foreskin fibroblasts, and wherein said foreskin fibroblast conditioned medium comprises at least 4 ng/ml of added basic fibroblast growth factor (bFGF), and being capable of maintaining said mammalian pluripotent stem cells in an undifferentiated, proliferative state through at least 87 passages, thereby maintaining the mammalian pluripotent stem cells in the undifferentiated state.
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10. A cell culture comprising:
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(i) mammalian pluripotent stem cells, wherein said mammalian pluripotent stem cells are capable of forming teratomas containing cells of the endoderm, mesoderm and ectoderm germ layers; and (ii) human foreskin fibroblast conditioned medium, wherein said foreskin fibroblast conditioned medium is of non-genetically modified human foreskin fibroblasts, and wherein said foreskin fibroblast conditioned medium comprises at least 4 ng/ml of added basic fibroblast growth factor (bFGF) and being capable of maintaining said mammalian pluripotent stem cells in an undifferentiated, proliferative state least 87 passages. - View Dependent Claims (11, 12, 13, 14, 15, 16)
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17. A method of producing a conditioned medium suitable for maintaining mammalian pluripotent stem cells in an undifferentiated state, the method comprising:
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(a) culturing non-genetically modified human foreskin fibroblast cells in a growth medium which comprises added basic fibroblast growth factor (bFGF) under conditions suitable for producing the conditioned medium; and (b) collecting said growth medium to thereby produce the conditioned medium, and (c) validating the ability of the conditioned medium to maintain said mammalian pluripotent stem cells in an undifferentiated state following step (b), thereby producing the conditioned medium suitable for maintaining the mammalian pluripotent stem cells in an undifferentiated state. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24, 25)
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26. A foreskin fibroblast conditioned medium which comprises at least 4 ng/ml added basic fibroblast growth factor (bFGF), wherein said foreskin fibroblast conditioned medium is of non-genetically modified human foreskin fibroblasts, wherein said foreskin fibroblast conditioned medium is suitable for maintaining mammalian stem cells in an undifferentiated state through at least 87 passages.
Specification