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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 in the separation of rat hepatocytes, viability, primary culture and MTT assay. Methods: Rat hepatocytes were isolated by collagenase and trypsin respectively. Cell viability was measured by trypan blue exclusion test and counted. The isolated hepatocytes were primary cultured and the morphological changes of hepatocytes were observed. MTT Test the proliferation of hepatocytes. Results: The number of hepatocytes isolated by collagenase was 2.4 times that of trypsin, and the activity of hepatocytes isolated by trypsin was 2.8 times that of collagenase. The hepatocytes isolated by collagenase were slow-growing and grew slowly Poor, and pancreatic enzyme isolated hepatocytes adherent fast, good growth; MTT test results show that pancreatic enzyme activity was significantly higher than that of collagenase. CONCLUSION: The number and purity of collagenase isolated cells are superior to those of trypsin, while the activity of pancreatic enzyme isolated cells is superior to that of collagenase.