Development and functional characterization of extrahepatic cholangiocyte lines from normal rats. Issue 11 (November 2015)

• Record Type: Journal Article
• 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
• Is Part Of: Digestive and liver disease. Volume 47:Issue 11(2015)
• Page Start: 964
• Page End: 972
• Publication Date: 2015-11
• Subjects: Digestive organs -- Diseases -- Periodicals; Liver -- Diseases -- Periodicals; 616.33005
• DOI: 10.1016/j.dld.2015.07.012
• Languages: English
• ISSNs: 1590-8658

Abstract

<abstract abstract-type="author" id="abs0005"> <title id="sect0005">Abstract</title> <sec> <title id="sect0010">Background</title> <p id="spar0005">Since limited <italic>in vitro</italic> tools exist for evaluating the pathophysiology of extrahepatic bile ducts, we aim to develop an extrahepatic cholangiocyte culture system from normal rats.</p> </sec> <sec> <title id="sect0015">Methods</title> <p id="spar0010">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 (SSTR<sub>2</sub>), 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.</p> </sec> <sec> <title id="sect0020">Results</title> <p id="spar0015">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 SSTR<sub>2</sub>, CFTR, AE2, VEGF-A and NGF. In extrahepatic cholangiocyte cultures, secretin increased cAMP (prevented by somatostatin), chloride efflux and proliferation.</p> </sec> <sec> <title id="sect0025">Conclusions</title> <p id="spar0020">Extrahepatic cholangiocyte cultures may be important for studying diseases targeting extrahepatic cholangiocytes such as biliary atresia.</p> </sec> </abstract>