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从创伤意外死亡的病人中获取肝组织,应用离体胶原酶Ⅳ灌注和消化的方法获得肝组织细胞悬液。根据差速离心将肝细胞和其它细胞进行分离,并根据枯否氏细胞和内皮细胞贴附时间的不同,分离出肝窦间隙内皮细胞,其存活率可达90%以上。肝细胞纯度可达95%。肝窦间隙内皮细胞经一周培养纯度可达90%以上。同时描述了人肝组织中肝细胞和肝窦间隙内皮细胞原代培养过程的形态特征,并对肝窦间隙内皮细胞进行了鉴定。
Hepatic tissues were harvested from patients who died of wounds accidentally. Hepatic tissue cell suspensions were obtained by perfusion and digestion of collagenase Ⅳ in vitro. Hepatic cells and other cells were separated by differential centrifugation, and the endothelial cells of hepatic sinusoidal space were separated according to the time of attachment of Kupffer cells and endothelial cells, and the survival rate was more than 90%. Liver cell purity up to 95%. Liver sinusoidal endothelial cells after one week of culture purity up to 90%. At the same time, the morphological characteristics of primary cultured endothelial cells of hepatocytes and hepatic sinusoids in human liver tissue were described, and the endothelial cells of hepatic sinusoidal space were identified.