LONG-TERM CULTURE AND CHARACTERISTICS OF NORMAL RAT-LIVER BILE-DUCT EPITHELIAL-CELLS

Background: A method was established for isolation and long-term culture of bile duct epithelial cells (BDEC) of normal adult rat liver that does not require the preparation of highly purified BDEC. Methods: After dissociation of the liver parenchyma by collagenase perfusion, the liver remnant containing the intact biliary tree was minced into small fragments, embedded in a rat tail collagen gel, and cultured for 6 days in hormonally defined serum-free Dulbecco's Modified Eagle Medium/F12 medium (SFDM). BDEC cultures were subsequently subcultured and maintained on rat tail collagen gels in SFDM medium containing 5 μmol/L forskolin and 5%-10% Nu Serum IV (Collaborative Research, Bedford, MA). Results: Established BDEC lines continued to express ductal specific markers including γ-glutamyl transpeptidase, cytokeratins 7 and 19, and a number of monoclonal antibody-defined bile duct antigens, such as OC.2, OC.3, and OV6. Conclusions: The availability of a method to establish normal BDEC lines will allow further investigation of the function of bile duct cells and their role in normal liver differentiation and carcinogenesis. © 1993.