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大鼠肝星状细胞的简易分离法
引用本文:朱欣彦,王胜兰,杨长青,杨文卓,张俊杰,孙龙娥.大鼠肝星状细胞的简易分离法[J].同济大学学报(医学版),2012,33(4):32-35.
作者姓名:朱欣彦  王胜兰  杨长青  杨文卓  张俊杰  孙龙娥
作者单位:同济大学附属同济医院消化内科,上海,200065
摘    要:目的在肝脏二步酶灌注法分离大鼠肝星状细胞的基础上,探讨一种简单易行且结果稳定的分离方法。方法采用大鼠肝脏二步酶灌注法对成年大鼠的肝脏进行消化,经密度梯度离心去杂质后得到肝星状细胞。台盼蓝拒染实验鉴定细胞活率,结蛋白(Desmin)免疫荧光鉴定肝星状细胞的纯度。结果肝星状细胞的得率为4.0×107以上,肝星状细胞纯度为96%左右,肝星状细胞存活率为(95.0±1.2)%。结论本实验采用的大鼠肝星状细胞的分离方法,结果稳定,细胞纯度较高,有利于进一步的生物学研究。

关 键 词:肝星状细胞  细胞分离  原代培养

A simple method for isolation of hepatic stellate cells from rat liver
ZHU Xin-yan,WANG Sheng-lan,YANG Chang-qing,YANG Wen-zhuo,ZHANG Jun-jie and SUN Long-e.A simple method for isolation of hepatic stellate cells from rat liver[J].Journal of Tongji University(Medical Science),2012,33(4):32-35.
Authors:ZHU Xin-yan  WANG Sheng-lan  YANG Chang-qing  YANG Wen-zhuo  ZHANG Jun-jie and SUN Long-e
Affiliation:(Dept.of Gastroenterology,Tongji Hospital,Tongji University,Shanghai 200065,China)
Abstract:Objective To develop a simple method for isolation of hepatic stellate cells (HSCs) from rat liver tissue. Methods The hepatocytes were obtained from digestion of adult Wistar rat liver by infusion of pronase and collagenase via portal vein; then the HSCs were isolated by density gradient centrifugation. The cell viability was determined by trypan blue exclusion test. The purity of HSCs was identified by immunofluorescence staining. Results The preparation yielded 4.0 X 107 cells, and the centrifugal elutriation resulted in pure HSC preparations (96% ) . The survival rate of hepatic stellate cells was (95.0±1.2) % . Conclusion A simple and efficient method for isolation of hepatic stellate cells from rat liver tissue has been developed in this study.
Keywords:hepatic stellate cell  cell isolation  primary culture
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