FGF having enhanced stability
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.
1. 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 fully defined culture medium comprising BMP4 and an FGF comprising the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4 until the cultured cell expresses mesoderm lineage markers.
2. The method of claim 1 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:3.
3. The method of claim 2 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:3.
4. The method of claim 1 , wherein the amino acid sequence of the FGF comprises the amino acid sequence of SEQ ID NO:4.
5. A method for reprogramming a human somatic cell into a human pluripotent stem cell, comprising:
culturing a somatic cell expressing a plurality of reprogramming factors sufficient to induce pluripotency in a fully defined culture medium comprising factors that support pluripotency of human pluripotent stem cells until the cell expresses markers indicative of a human pluripotent stem cell, wherein the medium comprises water, salts, amino acids, vitamins, glucose, insulin, selenium and an FGF comprising the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4.
6. The method of claim 5 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:3.
7. The method of claim 6 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:3.
8. The method of claim 5 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:4.
9. The method of claim 8 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:4.
10. The method of claim 5 , wherein the medium further comprises heparin.