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Abstract

BACKGROUND:

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.

OBJECTIVE:

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

METHODS:

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%

METHODS:

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.

RESULTS AND CONCLUSION:

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.

Funding:

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

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