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 culturing human pluripotent stem cells, comprising the step of culturing a human pluripotent stem cell in a fully defined culture medium comprising factors that support pluripotency of human pluripotent stem cells, wherein the culture medium comprises an FGF comprising the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4.
2. The method of claim 1 , wherein the human pluripotent stem cells are selected from the group consisting of human embryonic stem cells and human induced pluripotent stem cells.
3. The method of claim 1 , wherein the medium further comprises heparin.
4. The method of claim 1 , wherein the FGF is provided in the culture medium at a concentration of less than 40 ng/ml.
5. The method of claim 1 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:3.
6. The method of claim 5 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:3.
7. The method of claim 1 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:4.
8. The method of claim 7 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:4.
9. A fully-defined culture medium comprising:
(i) an FGF comprising the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4;
(ii) insulin or an insulin-like growth factor (IGF); and
(iii) TGFβ or Nodal;
wherein the culture medium supports culture of human pluripotent stem cells in an undifferentiated state.
10. The medium of claim 9 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:3.
11. The medium of claim 10 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:3.
12. The medium of claim 10 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:4.
13. The medium of claim 12 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:4.
14. The medium of claim 9 , wherein the FGF is provided in the medium at a concentration of less than 40 ng/ml.
15. A composition comprising:
a human pluripotent stem cell and a fully defined culture medium comprising
(i) an FGF comprising the amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4;
(ii) insulin or an IGF; and
(iii) TGFβ or Nodal; and
wherein the fully defined culture medium maintains the human pluripotent cell in an undifferentiated state.
16. The composition of claim 15 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:3.
17. The composition of claim 16 , wherein the FGF comprises the amino acid sequence of SEQ ID NO:4.
18. The composition of claim 15 , wherein the FGF is provided in the medium at a concentration of less than 40 ng/ml.
19. The composition of claim 15 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:3.
20. The composition of claim 15 , wherein the amino acid sequence of the FGF consists of the amino acid sequence of SEQ ID NO:4.
21. The method of claim 1 , wherein the culture medium is albumin-free.
22. The composition of claim 9 , wherein the culture medium is albumin-free.
23. The composition of claim 15 , wherein the culture medium is albumin-free.