DIFFERENTIATION OF BONE MARROW MESENCHYMAL STEM CELLS TO CARDIOMYOCYTES IN VITRO / 解剖学报

ABSTRACT

Objective To establish a new cell resource of stem cell transplantation for heart failure therapy,by using cardiomyocytes from rat marrow mesenchymal stem cells(MSCs). Methods MSCs were isolated and cultured from rat bone marrow,then induced by 5aza(5,10,15,20 ?mol/L) for 24 hours.The cultured cells were observed with phase-contrast microscopy.The immunohistochemical technique and laser scanning confocal microscopy(LSCM) were applied for revealing the expressions of desmin,?-sarcomeric actin and C-TnT.The induced cells were evaluated by a transmission electron microscope.GATA4 and(?-MHC) expressions were detected by relative quantitative RT-PCR after 7,21,28 days of inducement respectively. Results MSCs induced in vitro by 5-aza could be identified by the positive staining for desmin,?-sarcomeric actin and C-TnT.The number of cells stained positively in the 5 ?mol/L was greater than that of other groups.Transmission electron microscope showed that paralleled myofilaments were formed.RT-PCR assessment showed that the differentiated cells began to express GATA4 from day 7 to day 28 of differentiation and began to express ?MHC from day 21 to day 28 of differentiation.Conclusion Cardiomyocyte-like cells can be generated from MSCs in vitro and the marrow mesenchymal stem cells are one of the best resources of donor cardiomyocytes.