Abstract: In order to build method of a stable isolation and culture of adult myocytes, adult rat myocytes were isolated with bioenzymes (including 5 g/L trypsin and 1 g/L colleagase) perfusing tissue through acceding aorta of the heart. Calcium-tolerant myocytes were harvested, purifyed and cultured with DMEM culture medium . The myocytes quality was evaluated with morphology and trypan blue dye. States of myocytes growing in culture were observed with optical microscope and electron microscope photography. Result: Survival rate of aduld rat myocytes was 91.7%, survival myocytes were motionlessly attached to the bottom of culture plates, they were intact and rod-shaped, their length/width ratio was 4～6: 1. Conclusion: The method of isolation and culture of adult rat myocytes was successful.

Key words: adult rat myocytes, trypsin, colleagase, separation technique