Electrical stimulation promotes the differentiation of rat bone marrow mesenchymal stem cells to cardiomyocyte induced by 5-azacytidine in vitro

Objective To investigate the effects of electrical stimulation on the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to cardiomyocyte induced by 5-azacytidine in vitro. Methods MSCs from Sprague-Dawley (SD) rats were cultured and passed repeatedly to P3. MSCs were treated with 5-azacytidine (10 μmol/L) and incubated for 24 h.The induced MSCs were divided to stimulated group and non-stimulated group, and every groups divided by incubated for 1,2,3 and 4 weeks were named as subgroup Ⅰ,Ⅱ, Ⅲ and Ⅳ respectively.MSCs of stimulated group were stimulated for 30 min every day by supra-threshold square biphasic pulses (2 ms duration, 1.5 Hz, 20 μA), and the stimulation was initiated 1 d after inducing. Light and electronic microscope were used to identify the influences of characteristic morphological of MSCs in every subgroups,and immunocytochemistry was used to identify the expression of α-actin,cTnT, Cx43 in MSCs. Results The growth of MSCs in stimulated group was better than that of non-stimulated group. MSCs of stimulated group exhibited differentiation into cardiomyocyte-like cell at 1 week after inducing, earlier than that of no-stimulated group (2 weeks). In stimulated subgroup Ⅰ, scattered myogenic structure was observed in the plasma of some cells under electronic microscope, and α-actin,cTnT expressed in some cells, but not that be observed in non-stimulated subgroup Ⅰ. In cells of stimulated subgroups Ⅱ to Ⅳ, the expression level of α-actin, cTnT, Cx43 of were all higher than that of non-stimulated subgroups respectively. Conclusion Electrical stimulation (simulating the heart beating) could redound differentiation of rat bone marrow mesenchymal stem cells (MSCs) to cardiomyocyte induced by 5-azacytidine in vitro.