Differentiation protocol for making human cardiomyocytes
First Claim
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1. A method of differentiating human embryonic stem (hES) cells into cardiomyocyte lineage cells, comprising:
- a) obtaining hES cells and forming an embryoid body in a cell culture;
b) culturing the embryoid body from a) in the cell culture with activin, transforming growth factor (TGF), and insulin-like growth factor (IGF),c) subsequently adding bone morphogenic protein (BMP), fibroblast growth factor (FGF), insulin, and platelet-derived growth factor (PDGF);
to the cell culture from b) and thend) adding a nucleotide analog that affects DNA methylation to the cell culture of c) thereby differentiating hES cells into cardiomyocytes.
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Abstract
This invention provides populations human cells of the cardiomyocyte lineage. The cells are obtained by causing cultures of pluripotent stem cells to differentiate in vitro, and then harvesting cells with certain phenotypic features. Differentiated cells bear cell surface and morphologic markers characteristic of cardiomyocytes, and a proportion of them undergo spontaneous periodic contraction. Highly enriched populations of cardiomyocytes and their replicating precursors can be obtained, suitable for use in a variety of applications, such as drug screening and therapy for cardiac disease.
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Citations
17 Claims
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1. A method of differentiating human embryonic stem (hES) cells into cardiomyocyte lineage cells, comprising:
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a) obtaining hES cells and forming an embryoid body in a cell culture; b) culturing the embryoid body from a) in the cell culture with activin, transforming growth factor (TGF), and insulin-like growth factor (IGF), c) subsequently adding bone morphogenic protein (BMP), fibroblast growth factor (FGF), insulin, and platelet-derived growth factor (PDGF);
to the cell culture from b) and thend) adding a nucleotide analog that affects DNA methylation to the cell culture of c) thereby differentiating hES cells into cardiomyocytes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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