Abstract： Objective To investigate the isolation and culture ,and osteogenic differentiation of rat muscle derived stem cells ,and to compare those with the conventional bone marrow mesenchymal stem cells. Methods The skeletal muscle tissues of rat lower limbs were obtained and isolated by collagenase digestion to culture muscle derived stem cells. The bone marrow tissues were obtained and isolated to culture bone marrow mesenchymal stem cells. After transformed to the second generation,the two kinds of stem cells were cultured from basic medium to induction medium,and were differentiated to osteoblasts. The proliferation of the two stem cells was determined by CCK?8 method. Mineralized nodules staining and RT?PCR test were used to compare the osteogenic differentiation of the two stem cells. Results Both of the muscle derived stem cells and the conventional bone marrow mesenchymal stem cells could be induced to osteogenic differentiation. Higher cell activity and more nodule formation of the muscle derived stem cells were determined by CCK?8 method and the mineralized nodules staining,respectively. The expression of ALP, osteocalcin and osteopontin was found in both of the two stem cells by RT?PCR test,whereas higher expression was found in muscle derived stem cells. Conclusions Muscle derived stem cells can be successfully isolated and cultured from rat skeletal muscle tissue,and well induced to osteogenic differentiation,which might be an ideal source of seed cells for tissue engineering,as an alternative to bone marrow mesenchymal stem cells.