Enrichment and Characterization of Clonogenic Epithelial Cells from Adult Rat Liver and Initiation of Epithelial Cell Strains

• Published in: In Vitro Cellular & Developmental Biology 1987-05, Vol.23 (5), p.339-348
• Main Authors: Kazunori Furukawa, Shimada, Tomiko, England, Patricia, Mochizuki, Yohichi, Williams, Gary M.
• Format: Article
• Language: English
• Subjects: Animal cells; Animals; Biological and medical sciences; Biotechnology; Cell cultures. Hybridization. Fusion; Cell Division; Cell Fractionation; Cell growth; Cell lines; Cells, Cultured; Centrifugation; Cultured cells; Endothelial cells; Epithelial Cells; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Hepatocytes; Histocytochemistry; Liver; Liver - cytology; Liver cells; Male; Methods. Procedures. Technologies; Microscopy, Phase-Contrast; Molecular and cellular biology; Rats; Rats, Inbred F344; Seeding
• ISSN: 0883-8364; 2327-431X; 1475-2689
• DOI: 10.1007/BF02620990

A highly efficient method is described for obtaining proliferative epithelial cells from adult rat livers for the reproducible establishment of liver epithelial cell strains. When cells were isolated from livers of 10- to 15-wk-old male Fischer 344 rats by a collagenase-perfusion method, collected by centrifugation at 50 xg for 5 min, and cultured in Williams' medium E containing fetal bovine serum and dexamethasone, colonies of epithelial cells different in size and morphology from hepatocytes were obtained. Sequential perfusion with collagenase and dispase yielded numerous epithelial cell colonies. When isolated cells were fractionated by differential centrifugation, the great majority of hepatocytes were sedimented at 50 xg for 1 min, whereas many non-hepatocytic cells remained in the supernatant and could be sedimented by a second centrifugation at 50 xg for 5 min. Culture of the two fractions revealed that almost all the epithelial cell colonies were derived from cells in the non-hepatocytic cell fraction. The epithelial cells were cytochemically negative for γ-glutamyl transpeptidase activity, whereas an increase in the activity was detected in hepatocytes with duration in culture. Ultrastructural characteristics of hepatocytes were not found in the cells of newly established cell strains. These results suggest that adult rat liver epithelial cells propagable in culture were derived from a cell type other than the hepatocyte.