The aim was to establish the differentiating model " in vitro " and observe the characteristics of human skeletal muscle cells (HSMC). This study developed an " in vitro " technique of preparing cultures of HSMC obtained from 8 normal subjects. Culture of HSMC was identified with immunohistochemistry, and the characteristics in cytobiology , cytobiochemistry and cytomorphology were observed. CK activity and protein synthesis were measured.The results showed that cultured HSMC could highly differentiate, and there was positive correlations between protein synthesis, CK specific activity and the differentiating potency of HSMC. It suggested that the in vitro differentiation process of HSMC was similar to its in vivo differentiation process.