Es Cell International Pte Ltd

Other

Robert Passier

Christine Lindsay Mummery

This invention provides a method for testing a factor for cardiogenicity which comprises differentiating human embryonic stem (hES) cells to cardiomyocytes in the presence and absence of the factor wherein the human embryonic stem (hES) cells are cultured under a serum free condition comprising co-culture in the presence of END-2 cells or serum-free extracellular medium therefrom, and measuring the differentiation in the presence and absence of the factor. This invention also provides a method for identifying a cardiogenic factor, which comprises differentiating human embryonic stem (hES) cells to cardiomyocytes in the presence or absence of the factor wherein the human embryonic stem (hES) cells are cultured under a serum free condition comprising co-culture in the presence of END-2 cells or serum-free extracellular medium therefrom, and identifying the factor that affects the differentiation of human embryonic stem (hES) cells to cardiomyocytes in the presence or absence of the factor.

Cardiomyocyte differentiation

The present invention provides a method of enhancing the efficiency of differentiation of hES cells into cardiomyocytes which method comprises incubating the cells under serum free conditions. The method typically includes providing cells that induce cardiomyocyte differentiation by cell to cell contact. Differentiation to cardiomyocytes can occur via two routes, namely by spontaneous differentiation and by induced differentiation. Without wishing to be bound by theory the present inventors hypothesize that, in the case of induced differentiation, END-2 cells, for instance, are needed for aggregation to cause local high cell densities and in inducing differentiation of nascent mesoderm. This second step could be enhanced in any human embryonic stem cell line leading to the prediction that it will work in lines other than hES. In cell lines that undergo spontaneous differentiation, it is hypothesized that local induction of embryoid bodies in endoderm occurs. Typically for induced differentiation this method will also comprise culturing the hES cell with a cell excreting at least one cardiomyocyte differentiation inducing factor or with an extracellular medium therefrom, under conditions that induce differentiation.

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Utrecht, Netherlands

Robert Passier