Definitive endoderm
Disclosed herein are cell cultures comprising definitive endoderm cells and methods of producing the same. Also disclosed herein are cell populations comprising substantially purified definitive endoderm cells as well as methods for enriching, isolating and purifying definitive endoderm cells from other cell types.
1. An in vitro cell culture comprising a medium human pluripotent cells and human definitive endoderm cells having been differentiated from said human pluripotent cells, wherein at least 1 human definitive endoderm cell is present for every 5 human pluripotent cells present in said cell culture, wherein said medium comprises a Wnt family member.
2. The cell culture of claim 1 , wherein at least 1 human definitive endoderm cell is present for every 2 human pluripotent cells present in said cell culture.
3. The cell culture of claim 1 , wherein at least 2 human definitive endoderm cells are present for every 1 human pluripotent cells present in said cell culture.
4. The cell culture of claim 1 , wherein at least 5 human definitive endoderm cells are present for every 1 human pluripotent cells present in said cell culture.
5. The cell culture of claim 1 , wherein at least 10 human definitive endoderm cells are present for every 1 human pluripotent cells present in said cell culture.
6. The cell culture of claim 1 , wherein said human pluripotent cell comprises an embryonic stem cell.
7. The cell culture of claim 1 , wherein said medium lacks B27.
8. The cell culture of claim 1 , wherein said medium lacks serum replacement.
9. The cell culture of claim 1 , wherein said medium comprises a growth factor of the Nodal/Activin subgroup of the TGFβ superfamily.
10. The cell culture of claim 9 , further comprising a growth factor selected from the group consisting of Nodal, Activin A, Activin B and combinations thereof.
11. The cell culture of claim 1 , wherein said medium comprises Nodal.
12. The cell culture of claim 1 , wherein said medium comprises Activin B.
13. The cell culture of claim 1 , which is substantially free of cells derived from the ectoderm lineage.
14. The cell culture of claim 1 , wherein said human cells are nonrecombinant cells.
15. The cell culture of claim 1 , wherein said human definitive endoderm cells adhere to a surface of a culture vessel.
16. The cell culture of claim 1 , wherein said medium comprises Activin A.
17. The cell culture of claim 1 , wherein said Wnt family member comprises Wnt3A.