Primary culture and characterization of human hyperplastic extrahepatic biliary epithelial cells

Abstract

Objective Establishing the primary culture of human hyperpiastic biliary epithelial cells (BECs). Methods Cells were isolated by digestion with collagenase followed by scraping of the mu-cosa of human dilated extrahepatic bile duct. The characteristic and purity of the epithelial cells in primary culture were confirmed by immunocytochemistry and immunofluorescence with antibodies against cytokeratin 19 (CK-19) ,E-cadherin,Vimentin,α-SMA and S100A4. Results The BECs primary culture expanded quickly after attachment;Greater than 95% of the cells were positive for biliary epithelium marker CK-19 and E-cadherin,but weak positive or negative for vimentin,a marker for mesenchymal cells;The BECs were also negative for α-SMA,but positive for the epithelial-to-mesenchymal transition (EMT) marker S100A4. Conclusion We present an economic and easy-performing method for the isolation and culture of human extrahepatic BECs. These results indicate that highly purified human hyperplasic extrahepatic BECs can be prepared successfully(>98% ). And these cells were found to undergo EMT. It is suggested that the EMT of BECs contributes to liver biliary fibrogenesis. Key words: Biliary epithelial cells; Epithelial-mesenchymal transition