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目的:改良心肌细胞原代培养的方法条件、培养纯度,提高心肌细胞收获率和存活率。方法:取出生24h内SD大鼠心室,剪碎,0.05%胰蛋白酶、0.8～0.1mg/mlⅡ型胶原酶分次消化心室组织,差速贴壁法和化学试剂抑制非心肌细胞生长,纯化心肌细胞。用免疫组织化学方法检测细胞纯度。结果:心肌细胞存活率大于96%,细胞纯度亦为95%以上且细胞活性好。结论:本方法培养的原代心肌细胞存活率和纯度都大大提高,培养方法和操作步骤都得以简化,是一种理想心肌细胞原代培养方法。
OBJECTIVE: To improve the method of primary culture of cardiomyocytes, culture purity, improve cardiomyocyte harvest rate and survival rate. Methods: Ventricular tissues were isolated from ventricular tissue of SD rats during 24 hours of birth, cut into pieces, 0.05% trypsin and 0.8 ~ 0.1 mg / ml collagenase Ⅱ. Differential cardioplegia and chemical reagents were used to inhibit the growth of non-cardiomyocytes. cell. Cell purity was measured by immunohistochemistry. Results: The survival rate of cardiomyocytes was more than 96%, the cell purity was above 95% and the cell activity was good. CONCLUSION: The survival rate and purity of primary cardiomyocytes cultured by this method are greatly increased. The culture methods and operation steps are simplified, which is an ideal cardiomyocyte primary culture method.