We describe a rapid and reproducible technique for establishing primary cultures of skeletal muscle cells from mouse origin. This method was aimed at avoiding extensive enzymatic proteolysis which is commonly used for preparation of primary skeletal muscle cultures. It relies on a Stomacher blender that allows a rapid and regular mechanical dissociation of muscle samples by repeated shocks. Cultures have been compared to those obtained by a modification of the method of Yaffé (1993) based on tryptic dissociation of rat muscle thighs. The time of preparation was reduced to 1 h and 15 min as compared to 4 h with the technique of Yaffé. Both cultures displayed similar morphologies and exhibited comparable myogenesis processes. Cellular yield, rate of myotube formation and myotube numbers were similar. The expression of myogenesis markers were identical as assessed by determination of acetylcholine receptor number, creatine kinase activity and level of myosin light chain.