昆明医学院学报2012，Vol.33Issue(6)：18-20,24,4.
建立一种改良方法分离大鼠肝星状细胞
Establishment of a New Procedure for Isolating Rat Hepatic Stellate Cells
摘要
Abstract
Objective To build an efficient procedure for isolating rat hepatic stellate cells. Methods Primary hepatic stellate cells ( HSC) were isolated from normal Sprague-Dawley ( SD) rats by infusion and combined digestion of pronase E and collagenase Ⅳ ex situ. Hepatic stellate cells were purified by density centrifugation with 12% Nycodenz. Autofluorescenes, desmin and smooth muscle actin ( α -SMA) immunofluorescence staining identified and assayed purity of HSC. HSC were activated by culture on uncoated plastic tissue culture dish and culture in a higher glucose Dulbecco s modified eagles medium ( DMEM) supplemented with 10% fetal calf serum under 37℃ contained 5% CO2 95% air incubater. Results The harvest rate of hepatic stellate cells was about 3.7 0.6 107 per rat, and the viability was more than 90%. Hepatic stellate cells could be activated by culture for more than 7 days. Conclusion This reformed method is more efficient to isolate hepatic stellate cell and by culture the hepatic stellate cells can be activated.关键词
分离/大鼠/肝星状细胞Key words
Isolation/ Rat/ Hepatic stellate cells分类
临床医学引用本文复制引用
李玉莲,宋正己,范红,彭伟,陈艳敏,万苹..建立一种改良方法分离大鼠肝星状细胞[J].昆明医学院学报,2012,33(6):18-20,24,4.基金项目
国家自然科学基金资助项目(30960454) （30960454）
云南省科技计划应用基础计划项目(2009ZC169M) （2009ZC169M）
