摘要
目的:改进大鼠肝星状细胞的分离、培养方法,提高其分离质量。方法:取成年雄性Wistar大鼠,用链蛋白酶、胶原酶体内门静脉灌注消化大鼠肝脏细胞,经Nycodenz密度梯度离心分离肝星状细胞。台盼蓝拒染实验鉴定细胞活力,Desmin免疫细胞化学染色鉴定细胞纯度。结果:肝星状细胞得率为2.8×107/肝。肝星状细胞存活率在90%以上。Desmin阳性细胞原代培养达90%以上,传代培养达95%以上。结论:改良大鼠肝星状细胞的分离、培养方法,有利于原代大鼠肝星状细胞的生物学研究。
Objective： To improve the method to isolate and culture hepatic stellate cells （HSCs）. Methods：HSCs were isolated by Nycodenz density gradient centrifugation after the hepatocytes obtained from adult male Wistar rats were digested with pronase and collagenase infused via portal vein, The cell viability was detemtined by trypan blue exclusion test. The purity of HSCs was identified by detecting Desmin staining. Results： The yield rate of HSCs was 2.8×10^7 per rat liver, and the cell viability was more than 95%, The percentage of desmin-positive cell was more than 90% in primary culture and more than 95% in passage culture. Conclusion： This improved method to isolate and culture HSC can be used in the biological study of primary culture of HSCs.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2006年第6期603-605,共3页
Journal of China Medical University
基金
辽宁省教育厅高等学校科研基金资助项目(05L452)
关键词
肝星状细胞
细胞分离
原代培养
大鼠
hepatic stellate cell
cell isolation
primary culture
rat
