DIFFERENTIATION OF HUMAN EMBRYONIC STEM CELLS
First Claim
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1. A population of cells expressing markers characteristic of the pancreatic endocrine lineage that co-express NKX6.1 and insulin and wherein less than 10% of the cells in the population express glucagon.
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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 expressing markers characteristic of the pancreatic endocrine lineage that co-express NKX6.1 and insulin and minimal amounts of glucagon.
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Citations
7 Claims
- 1. A population of cells expressing markers characteristic of the pancreatic endocrine lineage that co-express NKX6.1 and insulin and wherein less than 10% of the cells in the population express glucagon.
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7. A method to generate a population of cells expressing markers characteristic of the pancreatic endocrine lineage that co-express NKX6.1 and insulin and wherein less than 10% of the cells in the population express glucagon, the method comprising the steps of:
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a. Culturing pluripotent stem cells, b. Differentiating the pluripotent stem cells into cells expressing markers characteristic of the definitive endoderm lineage, c. Differentiating the cells expressing markers characteristic of the definitive endoderm lineage into cells expressing markers characteristic of the pancreatic endoderm lineage, and d. Differentiating the cells expressing markers characteristic of the pancreatic endoderm lineage into cells expressing markers characteristic of the pancreatic endocrine lineage that co-express NKX6.1 and insulin and minimal amounts of glucagon by culturing the cells expressing markers characteristic of the pancreatic endoderm lineage in a medium supplemented with a factor capable of inhibiting BMP, a TGFβ
receptor signaling inhibitor, and a protein kinase C activator.
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Specification