目的:探讨新生大鼠心肌细胞原代培养的方法,培养存活率高和纯度高的心肌细胞。方法:无菌条件下取出出生1～2 d的SD大鼠心脏,用2.5 g/L胰蛋白酶和2.0 g/LⅡ型胶原酶等量混合液在37℃条件下分次消化心肌组织。以差速贴壁并将时间延长至90 min纯化心肌细胞,48 h后换液。结果:将心肌细胞24 h贴壁生长,于2～3 d心肌细胞伸出伪足,形成细胞簇,同步搏动,频率约80～130次/min,存活率为(93.9±2.8)%,纯度为(91.9±2.2)%。结论:以本实验的方法培养的心肌细胞存活率高、纯度高,是一种较为理想的原代心肌细胞培养方法。 Objective: To explore the method of primary culture of neonatal rat cardiomyocytes and to culture cardiomyocytes with high survival rate and high purity. Methods: Hearts of SD rats born 1 to 2 days were removed under aseptic conditions. Cardiomyocytes were digested with equal volume of 2.5 g / L trypsin and 2.0 g / L collagenase II at 37 ℃. Myocardial cells were differentiated and the time was extended to 90 min, and the cells were changed after 48 h. RESULTS: Myocardial cells grew adherently 24 h later, pseudopodia were formed in cardiomyocytes from 2 to 3 days, and clusters of cells were formed. They were pulsed synchronously with a frequency of about 80-130 beats / min, a survival rate of (93.9 ± 2.8)%, a purity (91.9 ± 2.2)%. Conclusion: The cultured cardiomyocytes cultured by this method have high survival rate and high purity, which is an ideal primary culture method of cardiomyocytes.