To investigate the way for in vitro culture of cardiac myocytes derived neonatal rat with micro-carriers in rotary cell culture system (RCCS), 1~2days old neonatal rat cardiac muscle cells were isolated by sequential digestion and pre-plating methods. The isolated cardiac myocytes were seeded into Cytodex-3 microcarrier in RCCS and cultured for 7 days. Morphological changes of the cells on microcarrier were observed by light microscope and scan electron microscopy. Metabolic indexes, including specific consumption rate of glucose, specific production rate of lactate, lactate transform rate were primarily measured. The results show that cells adhered into cytodex-3 microcarrier 12 hours after seeding and formed cells groups.Metabolic activity of the cells on microcarrier was vigorous. The above results demonstrate that culture cardiac myocytes on microcarrier in RCCS is a effective and brief way.