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US 8,785,184 B2Granted
Differentiation of human embryonic stem cells
AssigneeJanssen Biotech, Inc.
InventorJean Xu
Life of this patent
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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 cells capable of producing insulin following transplantation into an animal.
Claims2
A method to differentiate a population of human pluripotent stem cells into a purified population of pancreatic endoderm cells that co-express PDX-1 and NKX-6.1, but do not express CDX-2 and NGN-3 comprising the steps of:
- (a) culturing human pluripotent stem cells,
- (b) differentiating the human pluripotent stem cells into definitive endoderm cells by treating the human pluripotent stem cells with a medium supplemented with a TGF-β receptor agonist,
- (c) differentiating the definitive endoderm cells into a population of pancreatic endoderm cells that co-express PDX-1 and NKX6.1, but do not express CDX2 and NGN3 by treating the definitive endoderm cells with a first medium supplemented with FGF7, followed by culturing the cells in a second medium supplemented with FGF7, a factor capable of inhibiting BMP, retinoic acid, and a hedgehog signaling pathway inhibitor, and a TGF-β receptor agonist selected from the group consisting of:
- activin B, TGFβ-I, TGFβ-II, GDF-8, and GDF-11, and
- (d) selecting cells which co-express PDX-1 and NKX6.1 and do not express CDX2 and NGN3 to obtain a purified population of pancreatic endoderm cells that co-express PDX-1 and NKX-6.1, but do not express CDX-2 and NGN-3.
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Dependencies are inferred from the claim text.