昆明医学院学报2011，Vol.32Issue(7)：22-25,4.
原代大鼠肝细胞分离与纯化方法
Isolation and Purification of Primary Rat Hepatocytes
摘要
Abstract
Objective To establish an improved, simplified and effective hepatocyte isolation technique in order to increase cell production and decrease the prime cost. Methods The improved simple perfusion system was used to isolate rat hepatocytes by a two-step perfusion with collagenase in situ. Hepatocytes were separated from the dissociative cells by low speed centrifugation (500 rpm, 3 min, 3 times ) , and discontinous gradient centrifugation method was applied to purify the hepatocytes. The hepatocyte yields and viabilities were measured. Results The viability of hepatocytes obtained by the two-step perfusion with collagenase in situ was improved from (48.38 ±6.97) % to (92.63 ±2.50) % (P<0.01), and the purity was over 95% , after applying a discontinuous gradient centrifugation method. Conclusions This new method is a perfect technique for isolated rat hepatocytes. It has many advantages, such as need simple equipment, easy manipulation, high yields, viabilities and purity.关键词
肝细胞/分离/Ⅳ型胶原酶/PercollKey words
Hepatocyte/ Isolation/ Type IV collagenase/Percoll分类
基础医学引用本文复制引用
戴虹,何贵清,黄学惠,陈一晖,王超秀,李武..原代大鼠肝细胞分离与纯化方法[J].昆明医学院学报,2011,32(7):22-25,4.基金项目
云南省教育厅科研基金资助项目(08C0131) （08C0131）
