Fgf having enhanced stability
AssigneeWisconsin Alumni Research Foundation
InventorsGuokai Chen, James A. Thomson
Life of this patent
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Abstract
Methods are provided that exploit thermostable FGF-1 proteins for support of human pluripotent stem cell cultures. Also provided are compositions containing thermostable FGF-1 for culturing of human pluripotent stem cells.
Claims34
A method for culturing human pluripotent stem cells, comprising the step of culturing a human pluripotent stem cell in a medium comprising at least one thermostable fibroblast growth factor-one (FGF-1) that comprises amino acid substitutions at positions corresponding to positions 40, 47, and 93 of wildtype truncated FGF-1 (SEQ ID NO:2), wherein the medium comprising the thermostable FGF-1 is characterized by an enhanced ability to support pluripotency relative to a medium comprising truncated wild type FGF-1 (SEQ ID NO:2).
A fully-defined medium suitable for culturing human pluripotent cells in an undifferentiated state, the defined medium comprising a thermostable fibroblast growth factor-one (FGF-1) that comprises amino acid substitutions at positions corresponding to positions 40, 47, and 93 of wildtype truncated FGF-1 (SEQ ID NO:2).
A composition comprising:
- a human pluripotent stem cell and a culture medium comprising a thermostable fibroblast growth factor-one (FGF-1) that comprises amino acid substitutions at positions corresponding to positions 40, 47, and 93 of wildtype truncated FGF-1 (SEQ ID NO:2), and wherein the culture medium comprising the thermostable FGF-2 is suitable for culturing the human pluripotent cell in an undifferentiated state, and is characterized by an enhanced ability to support pluripotency relative to a medium comprising a truncated wild type FGF-1 (SEQ ID NO:2).
A method for differentiating a human pluripotent stem cell into a mesoderm lineage cell, the method comprising the step of:
- culturing the human pluripotent stem cell in a medium until the cell expresses mesoderm lineage markers, wherein the medium comprises BMP4 and a thermostable fibroblast growth factor-one (FGF-1) that comprises amino acid substitutions at positions corresponding to positions 40, 47, and 93 of wildtype truncated FGF-1 (SEQ ID NO:2), and wherein the medium comprising the BMP4 and the thermostable FGF-1 is characterized by an enhanced ability to support differentiation of a human pluripotent stem cell into a mesoderm lineage cell relative to a medium comprising BMP4 and a truncated wild type FGF-1 (SEQ ID NO:2).
A method for reprogramming a human somatic cell into a human pluripotent cell, comprising:
- culturing the somatic cell in a medium until the cell expresses markers indicative of a human pluripotent cell, wherein the medium comprises water, salts, amino acids, vitamins, a carbon source, insulin, selenium and at least one thermostable fibroblast growth factor-one (FGF-1) that comprises amino acid substitutions at positions corresponding to positions 40, 47, and 93 of wildtype truncated FGF-1 (SEQ ID NO:2), and wherein the medium comprising the thermostable FGF-1 is characterized by an enhanced ability to support pluripotency relative to a medium comprising a truncated wild type FGF-1 (SEQ ID NO:2).
Dependencies are inferred from the claim text.