Thermostable FGF-2 mutant having enhanced stability
AssigneeWisconsin Alumni Research Foundation
InventorsGuokai Chen, James A. Thomson, Zhonggang Hou
Life of this patent
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Abstract
Thermostable FGF-2 proteins having enhanced ability to support human pluripotent stem cell cultures are provided. Also provided are methods and compositions utilizing thermostable FGF-2 proteins.
Claims26
An isolated fibroblast growth factor-two, FGF-2, polypeptide, as depicted in SEQ ID NO:
- 2, that differs from wild type FGF-2, as depicted in SEQ ID NO:
- 1, at amino acid position 128, wherein the isolated FGF-2 contains an asparagine submitted for lysine at position 128 (a K128N substitution).
A method for culturing human pluripotent stem cells, the method comprising the step of culturing a human pluripotent stem cell in a medium comprising a K128-substituted thermostable fibroblast growth factor-two, FGF-2, as depicted in SEQ ID NO:
- 2, that differs from wild type FGF-2, as depicted in SEQ ID NO:
- 1, at position 128 of the wild type FGF-2, wherein the thermostable FGF-2 contains an asparagine submitted for lysine at position 128 (a K128N substitution).
A fully defined culture medium suitable for culturing human pluripotent cells in an undifferentiated state, the medium comprising thermostable fibroblast growth factor-two, FGF-2, as depicted in SEQ ID NO:2, that differs from wild type FGF-2, as depicted in SEQ ID NO:
- 1, at amino acid 128 of the wild type FGF-2, wherein the thermostable FGF-2 contains an asparagine submitted for lysine at position 128 (a K128N substitution).
A composition comprising:
- a human pluripotent stem cell;
- a medium suitable for culturing the human pluripotent cell in an undifferentiated state;
- and
- a thermostable fibroblast growth factor-two, FGF-2, as depicted in SEQ ID NO:2, that differs from wild type FGF-2, as depicted in SEQ ID NO:
- 1, at amino acid 128 of the wild type FGF-2, wherein the thermostable FGF-2 contains an asparagine submitted for lysine at position 128 (a K128N substitution).
A method for culturing reprogrammed human somatic cells comprising:
- culturing reprogrammed human somatic cells in a medium until they express markers indicative of induced pluripotent stem cells, wherein the medium comprises water, salts, amino acids, vitamins, a carbon source, insulin, selenium and thermostable fibroblast growth factor-two, FGF-2, as depicted in SEQ ID NO:2, that differs from wild type FGF-2, as depicted in SEQ ID NO:
- 1, at amino acid 128 of the wild type FGF-2, wherein the thermostable FGF-2 contains an asparagine submitted for lysine at position 128 (a K128N substitution).
Dependencies are inferred from the claim text.