Cells expressing pluripotency markers and expressing markers characteristic of the definitive endoderm
First Claim
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1. A method for deriving a population of cells expressing pluripotency markers and expressing markers characteristic of the definitive endoderm lineage comprising the steps of:
- a. Obtaining a population of cells expressing markers HNF-3β
, GATA-4, Mixl1, CXCR4 and SOX-17, characteristic of the definitive endoderm lineage, andb. Culturing the population of cells under hypoxic conditions, on a tissue culture substrate that is not pre-treated with a protein or an extracellular matrix in a medium supplemented with Activin A and wnt-3A, and IGF-1 or insulin, transferrin and selenium, wherein the cultured cells express the markers of the definitive endoderm lineage and pluripotency markers.
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Abstract
The present invention is directed to pluripotent cells that can be readily expanded in culture on tissue culture substrate that is not pre-treated with protein or an extracellular matrix, and do not require a feeder cell line. The present invention also provides methods to derive the pluripotent cell line from human embryonic stem cells.
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Citations
16 Claims
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1. A method for deriving a population of cells expressing pluripotency markers and expressing markers characteristic of the definitive endoderm lineage comprising the steps of:
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a. Obtaining a population of cells expressing markers HNF-3β
, GATA-4, Mixl1, CXCR4 and SOX-17, characteristic of the definitive endoderm lineage, andb. Culturing the population of cells under hypoxic conditions, on a tissue culture substrate that is not pre-treated with a protein or an extracellular matrix in a medium supplemented with Activin A and wnt-3A, and IGF-1 or insulin, transferrin and selenium, wherein the cultured cells express the markers of the definitive endoderm lineage and pluripotency markers. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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Specification