Long-term culture and characteristics of normal rat liver bile duct epithelial cells.

Abstract

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. 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 mumol/L forskolin and 5%-10% Nu Serum IV (Collaborative Research, Bedford, MA). Established BDEC lines continued to express ductal specific markers including gamma-glutamyl transpeptidase, cytokeratins 7 and 19, and a number of monoclonal antibody-defined bile duct antigens, such as OC.2, OC.3, and OV6. 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.