Cardiomyocyte Cell Populations
First Claim
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1. A method of inducing differentiation of cardiac cells from embryonic stem (ES) cells comprising culturing ES cells under conditions sufficient for formation of embryoid bodies (EBs), culturing the EBs under conditions sufficient for differentiation to hemangioblast/pre-erythroid cells, and isolating and reaggregating the hemangioblast/pre-erythroid cells in the presence of activated Notch to provide cardiac progenitor cells.
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Abstract
The present invention provides methods for inducing the differentiation of cardiac progenitor cells and cell populations produced by the methods of the invention. The invention further provides a method of screening for agents that affect cardiomyocytes, and a method of cardiomyocyte replacement therapy.
9 Citations
28 Claims
- 1. A method of inducing differentiation of cardiac cells from embryonic stem (ES) cells comprising culturing ES cells under conditions sufficient for formation of embryoid bodies (EBs), culturing the EBs under conditions sufficient for differentiation to hemangioblast/pre-erythroid cells, and isolating and reaggregating the hemangioblast/pre-erythroid cells in the presence of activated Notch to provide cardiac progenitor cells.
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20. A method of inducing differentiation of cardiac cells from embryonic stem (ES) cells comprising culturing ES cells under conditions sufficient to form embryoid bodies (EBs), wherein the ES cells contain a nucleic acid encoding the active intracellular domain of Notch4 operably linked to a regulatory element that controls inducible expression by an inducer;
- culturing the EBs in serum for about 3 days;
isolating Flk-1+ cells;
reaggregating the Flk-1+ cells in the presence of the inducer for about 24 hours to provide cardiac progenitor cells; and
culturing said cardiac progenitor cells in the absence of the inducer in serum free medium to provide cardiomyocytes.
- culturing the EBs in serum for about 3 days;
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27. A method of inhibiting the differentiation of cardiac cells from embryonic stem (ES) cells comprising culturing ES cells under conditions sufficient for formation of erythroid bodies (EBs), culturing the EBs under conditions sufficient for differentiation to Bry+/Flk-1−
- cells, isolating Bry+/Flk-1−
cells and reaggregating the Bry+/Flk-1−
cells in the presence of an inhibitor of Notch. - View Dependent Claims (28)
- cells, isolating Bry+/Flk-1−
Specification