摘要
目的探讨体外原代培养大鼠肝细胞的分离方法。方法以Wistar大鼠做为肝细胞供体 ,分别采用直接分离法与胶原酶消化法分离获取肝细胞并进行原代培养 ;以台盼蓝染色法测细胞存活率 ,在位相差倒置显微镜下观察细胞形态变化 ,MTT法测培养细胞活性 ,并检测不同时期培养上清液中白蛋白的含量。结果直接分离法获取的肝细胞活性及功能欠佳 ,而胶原酶消化法所获取的肝细胞形态完整、贴壁良好、活性高、功能强。结论经胶原酶消化分离获取肝细胞的方法优于直接分离法。
ObjectiveTo explore the isolating methods of rat hepatocytes for primary culture.MethodsRat hepatocytes were isolated by direct insolation and collagenase digestion respectively and were primarily cultured in medium.The viability as assessed by typan blue exclusion and MTT method.The morphologic changes of cultured hepatocytes were observed and the concentrations of albumin in the supernatant in different cultural period were examined.ResultsHepatocytes got by the method of direct isolation were not good enough on the viability and function.The cells,however,got by collagenase digestion were intact. The viability and function were good.ConclusionThe method of collagenase digestion for the isolation of hepatocytes for primary culture is better than that of direct isolation.
出处
《河北医科大学学报》
CAS
2000年第4期199-201,共3页
Journal of Hebei Medical University
基金
硕士研究生课题分题
关键词
肝细胞
分离
提纯
胶原酶消化分离
原代培养
hepatocytes/isol
collagenase digestion/methods
hepatocytes/cytol
