Isolation and culture of biliary epithelial cells from the biliary tract fraction of normal rats

Abstract

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.