Direct differentiation method for making cardiomyocytes from human embryonic stem cells
First Claim
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1. A direct differentiation method of obtaining cardiomyocyte lineage cells from primate pluripotent stem (pPS) cells, comprising in the following order:
- a) obtaining undifferentiated pPS cells;
b) plating the undifferentiated pPS cells without forming embryoid bodies directly onto a solid surface comprising a substrate to which cardiomyocyte lineage cells adhere without forming embryoid bodies;
c) permitting the plated pPS cells to adhere onto the substrate;
d) culturing the adhered cells in the absence of serum or feeder cells, but in the presence of activin and a bone morphogenic protein to produce cardiomyocyte lineage cells;
e) harvesting the cardiomyocyte lineage cells from the culture.
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Abstract
This invention provides a new procedure for generating cardiomyocyte lineage cells from embryonic stem cells for use in regenerative medicine. Differentiating by way of embryoid body formation or in serum is no longer required. Instead, the stem cells are plated onto a solid substrate, and differentiated in the presence of select factors and morphogens. After enrichment for cells with the appropriate phenotype, the cells are allowed to cluster into cardiac bodies™, which are remarkably homogeneous and suitable for the treatment of heart disease.
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Citations
14 Claims
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1. A direct differentiation method of obtaining cardiomyocyte lineage cells from primate pluripotent stem (pPS) cells, comprising in the following order:
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a) obtaining undifferentiated pPS cells; b) plating the undifferentiated pPS cells without forming embryoid bodies directly onto a solid surface comprising a substrate to which cardiomyocyte lineage cells adhere without forming embryoid bodies; c) permitting the plated pPS cells to adhere onto the substrate; d) culturing the adhered cells in the absence of serum or feeder cells, but in the presence of activin and a bone morphogenic protein to produce cardiomyocyte lineage cells; e) harvesting the cardiomyocyte lineage cells from the culture. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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