比较两种消化酶对分离大鼠肝细胞的影响

Comparation of Two Digestive Enzymes on the Influence of Isolated Rat Hepatocytes

目的:比较胶原酶和胰蛋白酶在分离大鼠肝细胞数量、活力、原代培养及MTT试验等四个方面的差别。方法:分别用胶原酶和胰酶分离获取大鼠肝细胞,通过台盼蓝拒染试验测细胞活力并计数,将分离的肝细胞进行原代培养并观察肝细胞形态变化,采用噻唑蓝(MTT)试验测肝细胞的增殖状况。结果:胶原酶分离的肝细胞数量是胰酶的2．4倍,而胰酶分离的肝细胞活力是胶原酶的2．8倍;胶原酶分离的肝细胞原代培养时贴壁慢,生长不良,而胰酶分离的肝细胞贴壁快,生长良好;MTT试验结果表明胰酶分离培养的细胞活性明显高于胶原酶。结论:胶原酶分离的细胞数量及纯度优于胰酶,而胰酶分离的细胞活力优于胶原酶。

Objective： To compare the differences of collagenase and trypsin on isolated rat hepatocytes including quantity,viability, primarily culture and MTT experiment. Method： The hepatocytes was isolated and harvested by collagenase and trypsin from Wistar rats. The viability of hepatocytes was tested by trypan blue exclusion and the quantity was calculated at the same time. The morphological change of primarily cultured hepatocytes was observed. The proliferative hepatocytes was tested by MTT experiment. Result： The isolated hepatocytes quantity by eollagenase was 2.4 times higher than trypsin, but the isolated hepatocytes viability by trypsin was 2.8 times greater than collagenase. The isolated hepatocytes by collagenase grew slowly,but the isolated hepatocytes by trypsin grew fast. MTT experiment showed the activity of isolated hepatocytes by trypsin was greater than collagenase obviously. Conclusion： The quantity and purity of hepatocytes by eullagenase was superior than trypsin, but the viability of hepatocytes by trypsin was better than collagenase.