Abstract

Pluripotent embryonic stem cells (ESC) have the ability to differentiate into a variety of cell lineages in vitro, including cardiomyocytes. Successful applications of ESC-derived cardiomyocytes in cell therapy and tissue engineering were limited by difficulties in selecting the desired cells from the heterogeneous cell population. We describe a simple method to generate relatively pure cardiomyocytes from mouse ESCs. A construct comprising mouse cardiac α-myosin heavy chain (MHC) promoter driving the neomycin resistance gene and SV40 promoter driving the hygromycin resistant gene designated pMHC-neo/SV40-hygro, was stably transfected into mouse ESCs. The transgenic ESC line, designated MN6 retained the undifferentiated state and the potential of cardiogenic differentiation. After G418 selection, more than 99% of cells expressed α-sarcomeric actin. Immunocytological and ultrastructural analysis demonstrated that, the selected cardiomyocytes were highly differentiated. Our results represent a simple genetic manipulation used to product essentially pure cardiomyocytes from differentiating ESCs. It may facilitate the development of cell therapy in heart diseases.

Key words: Embryonic stem cells, α-myosin heavy chain promoter, cardiomyocytes, differentiation, genetic enrichment.

Abbreviation

ESC, Embryonic stem cells; EB, emybryoid bodies; MHC, myosin heavy chain; MEF, mouse embryonic fibroblasts; FBS, fetal bovin serum; AKP,alkaline phosphatase; LIF, leukemia inhibitor factor.