新生大鼠心肌细胞的原代培养及鉴定

Culture and identification of cardiomyocytes in neonatal rats

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。

AIM：To develop a method of primary culture of neonatal rat cardiomyocytes with high survival cell rate and high purity. METHODS： Hearts of 1 to 2dayold neonatal Sprague Dawley rats were removed under sterile conditions with 0.25% trypsin and 02% collagenase type II at 37℃ under the conditions of graded digestion myocardial tissue. Differential adherence purification was applied. The time was extended to 90 min and the medium was changed after 48 h. RESULTS： Adherent growth of myocardial cells was observed at 24 h and pseudopodia growth at 2-3 days. Myocardial cells formed cell clusters and the synchronous beat frequency was about 80-130 times/min with survival rate of （93.9±2.8）% and purity of （91.9±2.2）%. CONCLUSION： We developed a method to culture myocardial cells with high survival cell rate and high purity.