Kumar U, Jordan T W
Liver. 1986 Dec;6(6):369-78. doi: 10.1111/j.1600-0676.1986.tb00306.x.
A method has been developed for the isolation of a population of cells enriched in epithelial lining cells from the bile ducts of normal rats. The procedure utilized digestion by pronase of the white strands of biliary and connective tissue which remained after hepatocytes had been mechanically removed from collagenase-perfused liver. The resulting cell population was enriched in cells whose ultrastructure resembled that of the epithelial cells of intrahepatic bile ducts. Contamination with hepatocytes, hepatocyte nuclei and erythrocytes was less than 2%. The cells have been maintained in short-term culture. The major morphological change during the first 2 days of culture was proliferation of microvilli, but cell protein composition was unchanged when analysed by polyacrylamide gel electrophoresis. A rabbit antiserum against bovine hoof prekeratin was used to immunohistochemically stain the intermediate filaments of biliary epithelium and was shown to stain more than 90% of the cells in the isolated cell population.
已开发出一种从正常大鼠胆管中分离富含上皮衬里细胞群体的方法。该程序利用链霉蛋白酶消化在肝细胞从胶原酶灌注的肝脏中机械去除后残留的胆管白色条索状和结缔组织。所得细胞群体富含超微结构类似于肝内胆管上皮细胞的细胞。肝细胞、肝细胞核和红细胞的污染率低于2%。这些细胞已在短期培养中维持。培养的前两天主要的形态学变化是微绒毛的增殖，但通过聚丙烯酰胺凝胶电泳分析时细胞蛋白质组成未发生变化。使用兔抗牛蹄前角蛋白抗血清对胆管上皮的中间丝进行免疫组织化学染色，结果显示在分离的细胞群体中超过90%的细胞被染色。
