A Novel Experimental Study on Establish a Myoblasts Differentiated Cell Sheet Using Induced Adipose-Derived Stem Cell Technology

The efficient treatment of muscular disease represents one of the unsolved problems in clinical practice. The transplantation of myoblasts might improve muscle regeneration and function. The aim of this study was to fabricate myoblast differentiated cell sheets by induced stem cell technology. Canine Adipose-Derived Stem Cells (ADSCs) were isolated from fat tissue of groin and then cultured. There were two experimental groups: a myoblast differentiated group and a control group. ADSCs were cultured in cell culture medium (control group) and medium with 10 mu M 5-azacytidine plus 5% horse serum (myoblast differentiated group). The ADSCs were stimulated with 100 mu g/ml vitamin C for 3 days, 50 mu g/ml vitamin C for other 18 days to form cell sheet. Then, cell sheets were transplanted to the subcutaneous area of nude mice. The surface structure of cell sheet was observed under the scanning electronic microscope. Meanwhile, Quantitative Real-time PCR (qPCR), western blot and immunohistochemical analysis were performed to detect the expressions of myoblast markers, alpha-SMA, Desmin and PAX7. The result showed myogenic differentiation was observed in the cells cultured with 5-azacytidine, and myoblast-like cells were identified form the retrieved implants. This study indicates that adipose-derived stem cells could be differentiated into myoblast-like cells in a format of cell sheet. It may provide a practical approach to boost muscle regeneration.