Abstract
Culturing murine embryonic stem (ES) cells within embryoid bodies (EBs) has been reported to reproduce cardiomyocyte development from primitive precursor cells to highly specialized phenotypes of cardiac tissue. We show here that the specific inhibitor of phosphatidylinositol-3-kinase (PI-3-kinase), LY294002, blocks the growth and induces apoptosis as well as necrosis of D3 ES cells within early EBs. Treatment of EBs from day 3 to day 7 with 50 μM LY294002 resulted in a massive loss of α-actinin-stained cardiomyocytes after plating the EBs for additional 7 days. In parallel we observed a strong decrease in the number of EBs containing area(s) with beating cardiomyocytes. The specific action of the PI-3-kinase inhibitor on development of cardiomyocytes was demonstrated by the observation that formation of endothelial cells was not affected in the same EBs. Our results provide the first evidence that signal transduction via the PI-3-kinase pathway is essential for mammalian early cardiomyocyte development.
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