Cardiomyocytes and methods of producing and purifying cardiomyocytes
AssigneeCalifornia Stem Cell, Inc.
InventorGABRIEL NISTOR
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Abstract
The invention provides methods for producing a culture of cardiomyocytes and cultures of cardiomyocytes. Exemplary methods of producing and cultures of cardiomyocytes include a population of cells including cells having spontaneous and periodic electrical activity, and/or including nodal, sino-atrial or pacemaker cells; immature cardiomyocytes (cardiomyoblasts); mature contractile cardiomyocytes; or a mixed population of two or more of such cells.
Claims13
A method for producing a culture of cardiomyocytes, comprising:
- A) Providing a culture of stem cells that are at least 90% confluent or the cells have overgrown to form multiple layers of cells, or proliferating stem cells until the cells are at least 90% confluent or the cells have overgrown to form multiple layers of cells;
- and
- B) Inducing formation of mesoderm by contacting the overgrown stem cells with a bone morphogenic protein (BMP) receptor ligand and an fibroblast growth factor (FGF) receptor ligand for a period of about 2 to 15 days;
- C) promoting cardiomyocyte formation by contacting mesoderm cells with neural cells or endoderm cells, or a neural cell or endoderm cell conditioned culture supernatant, for a period of about 1 to 21 days, thereby producing a culture of cardiomyocytes.
2-69. (canceled)
(canceled)
74-80. (canceled)
A cell culture of immature cardiomyocytes (cardiomyblasts), wherein 50% or more of said culture comprises immature cardiomyocyte (cardiomyblast) cells, and wherein no more than about 30% of said cells beat or contract.
83-90. (canceled)
A cell culture of immature cardiomyocytes (cardiomyblasts), wherein 50% or more of said cell culture comprises immature cardiomyocyte (cardiomyblast) cells, and wherein 75% or more of said immature cardiomyocyte (cardiomyblast) cells survive anaerobic conditions for 30, 60 or more minutes.
A cell culture of immature cardiomyocytes (cardiomyblasts), wherein 50% or more of said culture comprises immature cardiomyocyte (cardiomyblast) cells, and wherein no more than about 25% of said immature cardiomyocyte (cardiomyblast) cells die when subjected to anaerobic conditions for 30, 60 or more minutes.
93-102. (canceled)
Dependencies are inferred from the claim text.