消化酶浓度和消化时间对幼鼠心肌细胞培养的影响

INFLUENCE OF DIGESTIVE ENZYME AND DIGESTION TIME ON MYOCARDIAL CELL CULTURE IN YOUNG RATS

目的观察不同浓度消化酶和消化时间对幼鼠心肌细胞培养的影响。方法用0.08g/L和0.25g/L胰蛋白酶液消化幼鼠心肌组织,时间分别为5、8、15min,用差时贴壁法分离、提纯、收取细胞进行培养,倒置显微镜下观察细胞形态和活力。结果 0.08g/L胰蛋白酶液消化8min的心肌细胞第2天出现明显的搏动现象,第3～4天细胞伸出伪足相互接触交织成网,逐渐形成细胞簇或细胞单层,搏动呈同步性,收缩明显有力,搏动频率为120～180min-1;而经其他浓度胰蛋白酶和消化时间处理得到的心肌细胞形态和活力均不理想。结论选择适宜的消化酶浓度和消化时间可获得形态、活力良好的心肌细胞。

Objective To observe the effect of different concentrations of digestive enzyme and different digestion time on culture of myocardial cells in young rats.Methods Using 0.08 g/L and 0.25 g/L trypsin to digest the cardiac tissue for 5,8 and 15 min,respectively.The cells were then separated,purified,and collected.The morphology and vitality of the cells were observed with inverted microscopy.Results The cardiac cells treated with 0.08 g/L trypsin and digested for 8 min.appeared pulsation on the second day,and at days 3-4,pseudopods stretched out and contacted each other,forming a network.The cell cluster or cell monolayer was formed gradually,showing a synchronic beating with a strong contraction,and pulsatory frequency of120-180 /min-1.The morphology and vitality of cardiac cells treated with other concentrations of trypsin and digestion time were unsatisfactory.Conclusion Cardiac cells with favourable morphology and vitality can be obtained by choosing an appropriate concentration of digestive enzyme and digestion time.