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Mesenchymal stem cells can differentiate into cardiomyocytes in vitro induced by different methods, such as chemical drug induction, autologous transplantation in vivo, and in vitro simulation of cardiac-like microenvironment, but these inducible methods show low induction rate, complex operation, and toxic side effects.


To investigate the role of H9C2 cell culture medium in the differentiation of mesenchymal stem cells into cardiomyocyte-like cells.


Mesenchymal stem cells were obtained by the whole bone marrow adherent culture and H9C2 cell culture medium was prepared as a culture medium. Then mesenchymal stem cells were co-cultured with H9C2 cell culture medium for 1, 3, 5, 7 days. H9C2 cells cultured in 10% Dulbecco ‘s modified Eagle's medium/Ham's nutrient mixture F-12 served as positive control groups, while mesenchymal stem cells cultured in 10%


Dulbecco's modified Eagle ‘s medium/Ham's nutrient mixture F-12 as negative control group. Immunofluorescence and western blot assay were used to detect expression of myocardial cell junction protein (desmin) and troponin T. Real-time quantitative PCR was applied to detect mRNA expression of myocardial cell trait genes, α-cardiac myosin heavy chain and β-myosin heavy chain.


After co-culture of H9C2 cell culture medium and mesenchymal stem cells for 7 days, the troponin T positive cells were up to (16 ±7)%, which was significantly higher than that of non-induced mesenchymal stem cells. Compared with the negative control group, the expression of troponin T protein and desmin after induction were significantly increased (P < 0.05) by western-blot detection; real-time PCR showed that the mRNA expression of α-cardiac myosin heavy chain and β-myosin heavy chain in differentiated cells were both up-regulated (P < 0.05). These findings suggest that H9C2 cell culture medium may induce the differentiation of mesenchymal stem cells into cardiomyocyte-like cells.


the Medical Science and Technology Project of Chongqing Health Bureau, No. 2009-1-51

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