An improved and easy protocol for primary epithelial cell culture from atretic tissue in biliary atresia

摘要

Biliary atresia (BA) is a lethal disease of infancy with obscure etiology. Insight into the pathogenesis of this disorder is limited by lack of availability of adequate epithelial tissue. Primary culture of human biliary epithelium may help to provide material for diagnostic and research purposes. However, culture of these cells from atretic tissue is a challenging task. We aimed to develop a reliable and easier protocol for culture of human biliary epithelial cells from excised atretic extrahepatic bile duct. An explant culture was performed using tissue obtained from 30 children with diseases of biliary tract. The culture showed florid cell growth in less than 3 weeks. Epithelial nature and biliary origin of cultured cells was confirmed using pancytokeratin and cytokeratin -7 antibodies. The protocol showed 100% success rate as cells could be cultured in all 30 patients. Moreover, the cells remained viable for a duration of over 3 months in most of the cases. This easier culture technique is likely to have an impact on the study of biliary cell pathophysiology, particularly in BA.