大鼠肝星状细胞简便分离法及鉴定  

Simple Method for Isolation and Identification of Hepatic Stellate Cells from Rat Liver

作　　者：陈玲肖[1] 李兰兰[1] 马炬明[2] 王驹[2] 

机构地区：[1]浙江中医药大学第二临床医学院,浙江杭州310053 [2]解放军117医院,浙江杭州310013

出　　处：《辽宁中医药大学学报》2014年第2期125-128,共4页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：浙江省年度重大科技专项计划项目(2011C130291-1)

摘　　要：目的：介绍一种简便、经济、稳定的大鼠肝星状细胞（hepatic stellate cell，HSC）分离方法，为肝纤维化的研究提供细胞模型。方法：取符合标准的雄性SD大鼠，体重约400-450g，行肝脏门静脉插管，灌洗肝脏后，先后采用链酶蛋白酶、Ⅳ型胶原酶、DNA酶I灌注，消化肝脏制成细胞悬液，20％Nycodenz密度梯度离心分离得到原代HSC。Desmin及α—SMA抗体免疫组化鉴定纯度。结果：HSC得率2×10^7／每肝，细胞存活率及纯度达95％以上。结论：该法简便、经济，细胞得率和纯度稳定，是一种实用的大鼠HSC分离方法。Objective： To develop a simple and efficient method for isolation of rat hepatic stellate cells （HSC ） . Methods： Adult male SD rats were scarified and the needle was inserted into the hepatic portal vein and the liver was lavaged to remove the blood and prepare for the liver digestion. The liver was then removed from the site and separated with enzyme digestion to achieve the cell suspension. The cell suspension was transferred onto the top of 20% Nycodenz and went to density separation. A white membrane on the top of centrifugal liquid containing the HSC was recovered. The purity of HSC was identified by immunohistochemical staining of Desmin and ex -SMA. Results： The yield of HSC was 2× 10^7 per rat. The viability and purity of HSC was confirmed above 95%. Conclusions： This method has effectively simplified the procedure of HSC isolation and the cell harvested is qualified for the further study on HSC.

关 键 词：SD大鼠 肝星状细胞 细胞分离技术 原代培养 细胞活性鉴定 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学]