Chamber‐specific differentiation of Nkx2.5‐positive cardiac precursor cells from murine embryonic stem cells

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Abstract
SPECIFIC AIMSDiversification of cardiomyocytes is believed to occur at a very early stage in cardiac development; however, little is known regarding chamber-specific differentiation of embryonic stem (ES) cell-derived cardiomyocytes. As embryoid bodies (EBs) contain cardiomyocytes in different stages with different chamber specificities, it has been difficult to evaluate the diversification of ES cell-derived cardiomyocytes. To circumvent these problems, we have established an ES cell line [Nkx2.5 green fluorescent protein (GFP) ES] in which GFP is knocked into the Nkx2.5 locus. This study seeks to 1) establish a system to track the differentiation of Nkx2.5(+) cardiomyocytes purified from EBs; 2) determine if the Nkx2.5(+) cell lineage diversifies into various cardiac cell types, including sinoatrial (SA) node-, atrial-, or ventricular-type cells; and 3) test if retinoic acid (RA) affects chamber-specific differentiation of ES cell-derived cardiomyocytes.PRINCIPAL FINDINGS1. Purification and differentiat...