Differentiation of cardiomyocytes in floating embryoid bodies is comparable to fetal cardiomyocytes

Embryonic stem cells will cluster and differentiate into embryoid bodies, which can develop spontaneous rhythmic contractions. From these embryoid bodies, cardiomyocytes can be isolated based on density by a discontinuous Percoll gradient. These cardiomyocytes differentiate into ventricular myocytes, which is demonstrated by the expression of the ventricular specific isoform of the myosin light chain 2 gene, In this study the functional expression of ion channels was compared between fetal cardiomyocytes (in vivo) and stem cell derived cardiomyocytes (in vitro). Sodium and calcium currents together with transient potassium currents could be detected in early developmental stages (<day 14) both in vivo and in vitro. In the early stages, we found a limited number of cells expressing I-Kr and virtual absence of I-Ks. The characteristics and distribution of currents are similar in both cell types. The current characteristics were identical for ventricular compared to atrial or undifferentiated stem cell derived cardiomyocytes, despite differences in expression of regulatory myosin light chain proteins. The myocyte differentiation was verified in a limited number of cardiomyocytes following the patch clamp proccdure by immunocytochemistry. (C) 2000 Academic Press.