Differentiation of human embryonic stem cells
First Claim
1. An in vitro cell culture comprising an isolated population of cells and a culture medium suitable for differentiating pluripotent stem cells,wherein greater than 85% of the cells are definitive endoderm cells,wherein said population of cells is obtained by differentiating in vitro pluripotent stem cells,wherein the culture medium lacks serum and is supplemented with BSA, GDF-8, 14-Prop-2-en-1-yl-3,5,7,14,17,23,27-heptaazatetracyclo[19.3.1.1˜
- 2,6˜
0.1˜
8,12˜
]heptacosa-1(25),2(27),3,5,8(26),9,11,21,23-nonaen-16-one and a factor selected from the group consisting of insulin and from about 1 ng/ml to about 50 ng/ml of IGF-1, andwherein said definitive endoderm cells express CXCR4 and do not express CD9.
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0 Petitions
Accused Products
Abstract
The present invention provides methods to promote the differentiation of pluripotent stem cells into insulin producing cells. In particular, the present invention provides a method to produce a population of cells, wherein greater than 85% of the cells in the population express markers characteristic of the definitive endoderm lineage.
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Citations
17 Claims
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1. An in vitro cell culture comprising an isolated population of cells and a culture medium suitable for differentiating pluripotent stem cells,
wherein greater than 85% of the cells are definitive endoderm cells, wherein said population of cells is obtained by differentiating in vitro pluripotent stem cells, wherein the culture medium lacks serum and is supplemented with BSA, GDF-8, 14-Prop-2-en-1-yl-3,5,7,14,17,23,27-heptaazatetracyclo[19.3.1.1˜ - 2,6˜
0.1˜
8,12˜
]heptacosa-1(25),2(27),3,5,8(26),9,11,21,23-nonaen-16-one and a factor selected from the group consisting of insulin and from about 1 ng/ml to about 50 ng/ml of IGF-1, andwherein said definitive endoderm cells express CXCR4 and do not express CD9. - View Dependent Claims (2, 3, 4)
- 2,6˜
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5. A method for generating a population of cells wherein greater than 85% of the cells in the population are definitive endoderm cells, comprising the steps of:
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a. culturing a population of pluripotent stem cells; and b. differentiating the population of pluripotent stem cells to a population of cells wherein greater than 85% of the cells in the population are definitive endoderm cells in medium lacking serum and supplemented with BSA, GDF-8, 14-Prop-2-en-1-yl-3,5,7,14,17,23,27-heptaazatetracyclo[19.3.1.1˜
2,6˜
0.1˜
8,12˜
]heptacosa-1(25),2(27),3,5,8(26),9,11,21,23-nonaen-16-one and a factor selected from the group consisting of insulin and from about 1 ng/ml to about 50 ng/ml of IGF-1. - View Dependent Claims (6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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16. An in vitro culture comprising an isolated population of cells in which greater than 85% of the cells are definitive endoderm cells and a cell culture medium,
wherein said population of cells is obtained by differentiating pluripotent stem cells into definitive endoderm cells, and wherein the cell culture medium lacks serum and is supplemented with BSA, GDF-8, 14- and Prop-2-en-1-yl-3,5,7,14,17,23,27-heptaazatetracyclo[19.3.1.1˜ - 2,6˜
0.1-8,12˜
]heptacosa-1(25),2(27),3,5,8(26),9,11,21,23-nonaen-16-one and either insulin or from about 1 ng/ml to about 50 ng/ml of IGF-1. - View Dependent Claims (17)
- 2,6˜
Specification