Isolation and osteogenic differentiation of skeletal muscle-derived stem cells for bone tissue engineering

Abstract

The purpose of this study was to investigate the isolation and culture of muscle-derived stem cells (MDSCs) and their capability to differentiate into osteoblasts in vitro. Skeletal muscle tissue was obtained from double hind limbs of New Zealand white rabbits under sterile conditions and isolated by collagenase digestion. Following passages in basic medium, the primary cells were desmin (+), myosin (+) and CD105 (+). Differentiation of MDSCs was induced by osteogenic medium. Using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, the differentiated cell population was found to proliferate faster than the undifferentiated. Alkaline phosphatase staining and alizarin red staining revealed that the differentiated cells were mineralized in vitro. Quantitative polymerase chain reaction assays also showed increased mRNA expression of osteogenic genes in differentiated cells. In conclusion, stem cells were successfully isolated and cultured from rabbit skeletal muscle tissue and were able to differentiate into osteoblasts following induction. These observations may indicate an ideal stem cell source for tissue engineering.