Differentiation of Human Embryonic Stem Cells
First Claim
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1. A method to differentiate a population of pluripotent stem cells into a population of cells expressing markers characteristic of the pancreatic endoderm lineage that co-express PDX-1, NKX-6.1, but do not express CDX-2 and NGN-3 comprising the steps of:
- a. Culturing the pluripotent stem cells,b. Differentiating the pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage, andc. Differentiating the cells expressing markers characteristic of the definitive endoderm lineage into cells expressing markers characteristic of the pancreatic endoderm lineage that co-express PDX1, NKX6.1, but do not express CDX2 and NGN3 by treating cells expressing markers characteristic of the definitive endoderm lineage 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, activin A, retinoic acid, and a hedgehog signaling pathway inhibitor.
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Abstract
The present invention provides methods to promote the differentiation of pluripotent stem cells into cells expressing markers characteristic of the pancreatic endocrine lineage that co-express PDX1, NKX6.1, but do not express CDX2 and NGN3.
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1 Claim
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1. A method to differentiate a population of pluripotent stem cells into a population of cells expressing markers characteristic of the pancreatic endoderm lineage that co-express PDX-1, NKX-6.1, but do not express CDX-2 and NGN-3 comprising the steps of:
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a. Culturing the pluripotent stem cells, b. Differentiating the pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage, and c. Differentiating the cells expressing markers characteristic of the definitive endoderm lineage into cells expressing markers characteristic of the pancreatic endoderm lineage that co-express PDX1, NKX6.1, but do not express CDX2 and NGN3 by treating cells expressing markers characteristic of the definitive endoderm lineage 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, activin A, retinoic acid, and a hedgehog signaling pathway inhibitor.
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