A RAPID PREPARATION OF PRIMARY CULTURES OF MOUSE SKELETAL-MUSCLE CELLS

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