Heuff G, Meyer S, Beelen R H
Department of Surgical Oncology, University Hospital, Free University, Amsterdam, Netherlands.
J Immunol Methods. 1994 Sep 14;174(1-2):61-5. doi: 10.1016/0022-1759(94)90010-8.
A new rapid method is described for the isolation and purification of rat and human Kupffer cells without the need of liver perfusion techniques. Rat livers or small human liver wedge biopsies obtained peroperatively were incubated with pronase under continuous pH registration. Kupffer cells were subsequently separated from other nonparenchymal cells by Nycodenz gradient centrifugation and purified by counterflow centrifugal elutriation. Identification of Kupffer cells was achieved on the basis of ultrastructural analyses and immunophenotyping.
本文描述了一种新的快速方法，用于分离和纯化大鼠和人类的库普弗细胞，无需肝脏灌注技术。术中获取的大鼠肝脏或人类小肝楔形活检组织在连续pH记录下用链霉蛋白酶孵育。随后通过 Nycodenz 梯度离心将库普弗细胞与其他非实质细胞分离，并通过逆流离心淘析进行纯化。基于超微结构分析和免疫表型鉴定库普弗细胞。
