Autologous serum enhances cardiomyocyte differentiation of rat bone marrow mesenchymal stem cells in the presence of transforming growth factor-beta 1 (TGF-beta 1)

摘要

In spite of previous reports, the role of transforming growth factor-beta 1 (TGF-beta 1) on cardiomyocyte differentiation, especially in the present autologous serum (AS) in culture medium, is still unclear. So, the purpose of this study was to investigate the potential of rat bone marrow mesenchymal stem cells (rBMSCs) to proliferate and differentiate towards cardiomyocyte lineage with the use of AS. Most expansion protocols use a medium supplemented with fetal bovine serum (FBS) as nutritional supplement. FBS is an adverse additive to cells that are proliferated for therapeutic purposes in humans because the use of FBS carries the risk of transmitting viral and bacterial infections and proteins that may initiate xenogeneic immune responses. Therefore, bone marrow cells were cultured in a medium supplemented with 10% AS, 10% FBS, and serum free medium (SFM). Then, rBMSCs were cultured with TGF-beta 1 (10 ng/ml) for 2 wk. The number of viable cells in AS and FBS groups were measured with MTT assay. Beating areas frequency, up to fourth week after plating, were monitored and evaluated daily. The characteristics of cardiomyocytes were assessed by semi-quantitative reverse transcription polymerase chain reaction and western blot. MTT result indicated that rBMSCs in AS proliferated markedly faster than FBS and SFM. The number of beating areas significantly increased in AS compared to FBS medium. A noticeable increase in the cardiac genes expression was observed in AS. Moreover, western blot analysis confirmed that cardiac proteins were increased in the AS condition. In conclusion, the present study could be extended toward the safe culture of MSCs for the treatment of heart defects.