Development and functional characterization of extrahepatic cholangiocyte lines from normal rats. Academic Article uri icon


  • BACKGROUND: Since limited in vitro tools exist for evaluating the pathophysiology of extrahepatic bile ducts, we aim to develop an extrahepatic cholangiocyte culture system from normal rats. METHODS: Extrahepatic ducts were dissected from rats, cut in half length-wise and cultured on collagen-I coated plates. Transepithelial electrical resistance was measured. At ∼85% confluence, in extrahepatic cholangiocytes we measured: (i) cell size and distribution, and expression for cytokeratin-19, secretin, secretin receptor and somatostatin receptor type II (SSTR2), cystic fibrosis transmembrane conductance regulator (CFTR), chloride bicarbonate anion exchanger 2 (AE2), vascular endothelial growth factor-A (VEGF-A) and nerve growth factor (NGF); and (ii) the effect of secretin and/or somatostatin on 3'-5'-cyclic adenosine monophosphate (cAMP) levels and proliferation. RESULTS: Cytokeratin-positive extrahepatic cholangiocytes were cultured for 6 passages to form a cell monolayer. Cholangiocytes proliferated to confluence over a 2-week period. The size of extrahepatic cholangiocytes averaged ∼16 μm. Extrahepatic ducts and cholangiocytes were positive for secretin, secretin receptor and SSTR2, CFTR, AE2, VEGF-A and NGF. In extrahepatic cholangiocyte cultures, secretin increased cAMP (prevented by somatostatin), chloride efflux and proliferation. CONCLUSIONS: Extrahepatic cholangiocyte cultures may be important for studying diseases targeting extrahepatic cholangiocytes such as biliary atresia.

published proceedings

  • Dig Liver Dis

author list (cited authors)

  • Venter, J., Francis, H., Meng, F., DeMorrow, S., Kennedy, L., Standeford, H., ... Alpini, G.

citation count

  • 12

complete list of authors

  • Venter, Julie||Francis, Heather||Meng, Fanyin||DeMorrow, Sharon||Kennedy, Lindsey||Standeford, Holly||Hargrove, Laura||Wu, Nan||Wan, Ying||Frampton, Gabriel||McMillin, Matthew||Marzioni, Marco||Gaudio, Eugenio||Onori, Paolo||Glaser, Shannon||Alpini, Gianfranco

publication date

  • January 2015