Janousek J, Strmen E, Gervais F
McGill Centre for the Study of Host Resistance, Montreal General Hospital Research Institute, Quebec, Canada.
J Immunol Methods. 1993 Aug 26;164(1):109-17. doi: 10.1016/0022-1759(93)90281-b.
Procedures to reproducibly obtain pure preparations of murine Kupffer cells are described. Pure Kupffer cell preparations obtained following collagenase digestion, metrizamide separation and centrifugal elutriation remained viable and maintained their phagocytic functions for at least 4-5 days in vitro. Furthermore, we determined the feasibility of extracting RNA from Kupffer cells obtained immediately following elutriation and after 2 and 4 days of culture in vitro. These RNA extracts were used to determine the level of cytokine gene expression in Kupffer cells.
本文描述了可重复获得小鼠库普弗细胞纯制剂的方法。通过胶原酶消化、甲泛葡胺分离和离心淘析获得的纯库普弗细胞制剂在体外至少4-5天内保持存活并维持其吞噬功能。此外,我们还确定了从淘析后立即获得的库普弗细胞以及体外培养2天和4天后获得的库普弗细胞中提取RNA的可行性。这些RNA提取物用于测定库普弗细胞中细胞因子基因表达水平。