| Objective: To optimizate the conditions of culturing mesenchymal stem cells in vitro; Utilization of 5–azacytidine differentiate meschymal stem cells into cardiocyte-like cells,providing a reference for follow-up experiments; Using myocardium lysate and supernatant of culturing myocardial cells , to discuss their ability to induce bone marrow mesenchymal stem cells into cardiomyocyte-like cells.Method: Isolation and culturing MSCs from rats'bone marrow on sterile conditions. Purified and cultured myoardial cells by differential adhesion method. Observed their biological characteristics under inverted microscope. Choose the third passaged MSCs well growing divided into three groups: I control group, culturing MSCs with High Glucose Dulbecco's modified Eagle's medium containing 10% Fetal bovine serum and no inducer. II Positive control group (5-aza induction group), With different concentrations of 5-aza role in MSCs for 24 hours, then culturing in High Glucose Dulbecco's modified Eagle's medium containing 10% Fetal bovine serum. III Supernatant induced group, Cardiac cell culture supernatants collected to prepare the conditioned medium to culture MSCs. IV Cardiac tissue lysates induced group, culture MSCs with cardiac tissue lasate. Changes of morphological characteristics of the MSCs that cultured for 4 weeks were observed. After the procedure of induction, hematoxylin and eosin staining were used to identify whether intercalated disk was existed while electron microscopys was used to assess the ultrstructure of the induced cells.Rusult: Supernatant was insufficient to induce myocardial cells of bone marrow MSCs to differentiate into cardiomyocyte-like cells. 5-aza, cardiac tissue lysates can induce bone marrow MSCs to differentiate into cardiomyocyte-like cells , self-beating with 90-110 times / min.Conclusion: The myocardium lysate can imitating the microenvironment of cardiocyte , induce bone marrow mesenchymal stem cells differentiated into myocardial-like tissue.
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