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 cultur
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.
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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 GDF-8 and a cyclic analine-pyridinotriazine comprising 5-Chloro-1,8,10,1
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 GDF-8 and a cyclic analine-pyridinotriazine comprising 5-Chloro-1,8,10,12,16,22,26,32-octaazapentacyclo[24.2.2.1˜3,7˜.1˜9,13˜.1˜14,18˜]tritriaconta-3(33),4,6,9(32),10,12,14(31),15,17-nonaen-23-one, 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 method increases the percentage of cells expressing CXCR4. 3. The method of claim 1, wherein the medium further comprises EGF, FGF or EGF and FGF and wherein the method enhances expression of SOX17. 4. The method of claim 1, wherein the medium is further lacking Wnt3a. 5. The method of claim 1, wherein the medium further comprises at least one factor selected from the group consisting of: PDGF-A, PDGF-B, PDGF-C, PDGF-D, VEGF, muscimol, PD98059, LY294002, U0124, U0126, and sodium butyrate. 6. A method of enhancing the differentiation of pluripotent stem cells into definitive endoderm lineage cells comprising differentiating pluripotent stem cells into definitive endoderm cells by culturing the pluripotent stem cells for about one to about three days with a medium lacking activin A, and containing GDF-8 and 5-Chloro-1,8,10,12,16,22,26,32-octaazapentacyclo[24.2.2.1˜3,7˜.1˜9,13˜.1˜14,18˜]tritriaconta-3(33),4,6,9(32),10,12,14(31),15,17-nonaen-23-one. 7. The method of claim 6, wherein the medium further comprises at least one factor selected from the group consisting of: EGF, FGF4, PDGF-A, PDGF-B, PDGF-C, PDGF-D, VEGF, muscimol, PD98059, LY294002, U0124, U0126, and sodium butyrate. 8. The method of claim 6, wherein the method comprises culturing the pluripotent stem cells with a medium lacking activin A, and containing GDF-8 and 5-Chloro-1,8,10,12,16,22,26,32-octaazapentacyclo[24.2.2.1˜3,7˜.1˜9,13˜.1˜14,18˜]tritriaconta-3(33),4,6,9(32),10,12,14(31),15,17-nonaen-23-one for about one day. 9. A method of enhancing the yield of definitive endoderm cells in a population comprising definitive endoderm cells comprising: differentiating a population of pluripotent stem cells into the population comprising definitive endoderm cells by culturing the pluripotent stem cells in a medium lacking activin A, and containing GDF-8 and 5-Chloro-1,8,10,12,16,22,26,32-octaazapentacyclo[24.2.2.1˜3,7˜.1˜9,13˜.1˜14,18˜]tritriaconta-3(33),4,6,9(32),10,12,14(31),15,17-nonaen-23-one. 10. The method of claim 9, wherein the method comprises culturing in the medium from about one to about three days. 11. The method of claim 9, wherein the medium further comprises at least one factor selected from the group consisting of: EGF, FGF4, PDGF-A, PDGF-B, PDGF-C, PDGF-D, VEGF, muscimol, PD98059, LY294002, U0124, U0126, and sodium butyrate. 12. The method of claim 1, wherein said method enhances differentiation into the definitive endoderm lineage when compared to cells treated with a medium comprising activin A. 13. The method of claim 1, wherein the cells are cultured in the medium for about one to seven days.
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