Differentiation of Pluripotent Stem Cells
The present invention is directed to methods to differentiate pluripotent stem cells. In particular, the present invention is directed to methods and compositions to differentiate pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage comprising culturing the pluripotent stem cells in medium comprising a sufficient amount of GDF-8 to cause the differentiation of the pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage.
1 . A method to differentiate pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage, comprising treating the pluripotent stem cells with a medium lacking activin A, and containing a compound selected from the group consisting of: EGF, FGF4, PDGF-A, PDGF-B, PDGF-C, PDGF-D, VEGF, GDF-8, muscimol, PD98059, LY294002, U0124, U0126, and sodium butyrate, for a period of time sufficient for the pluripotent stem cells to differentiate into cells expressing markers characteristic of the definitive endoderm lineage.
2 . The method of claim 1 , wherein the lacking activin A also contains at least one other compound selected from the group consisting of: an aniline-pyridinotriazine and a cyclic aniline-pyridinotriazine.