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Am J Physiol Gastrointest Liver Physiol 284: G1066-G1073, 2003. First published January 22, 2003; doi:10.1152/ajpgi.00260.2002
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Vol. 284, Issue 6, G1066-G1073, June 2003

Development and characterization of secretin-stimulated secretion of cultured rat cholangiocytes

Gianfranco Alpini1,4,5, Jo Lynne Phinizy3, Shannon Glaser3, Heather Francis3, Antonio Benedetti6, Luca Marucci6, and Gene LeSage1,2,7

Departments of 1 Internal Medicine and 2 Medical Biochemistry and Genetics, 3 Division of Research and Education, 4 Medical Physiology, Scott and White Hospital and Texas A&M University System, Health Science Center, College of Medicine and 5 Central Texas Veterans Health Care System, Temple 76504; 6 Department of Gastroenterology, University of Ancona, Ancona, Italy; and 7 University of Texas, Houston, Texas 77030

We sought to develop a cholangiocyte cell culture system that has preservation of receptors, transporters, and channels involved in secretin-induced secretion. Isolated bile duct fragments, obtained by enzyme perfusion of normal rat liver, were seeded on collagen and maintained in culture up to 18 wk. Cholangiocyte purity was assessed by staining for gamma -glutamyl transpeptidase (gamma -GT) and cytokeratin-19 (CK-19). We determined gene expression for secretin receptor (SR), cystic fibrosis transmembrane conductance regulator, Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchanger, secretin-stimulated cAMP synthesis, Cl-/HCO3 exchanger activity, secretin-stimulated Cl- efflux, and apical membrane-directed secretion in polarized cells grown on tissue culture inserts. Cultured cholangiocytes were all gamma -GT and CK-19 positive. The cells expressed SR and Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchanger, and secretin-stimulated cAMP synthesis, Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchanger activity, and Cl- efflux were similar to freshly isolated cholangiocytes. Forskolin (10-4 M) induced fluid accumulation in the apical chamber of tissue culture inserts. In conclusion, we have developed a novel cholangiocyte line that has persistent HCO<UP><SUB>3</SUB><SUP>−</SUP></UP>, Cl-, and fluid transport functions. This cell system should be useful to investigators who study cholangiocyte secretion.

bicarbonate secretion; bile flow; intrahepatic bile ducts; adenosine 3',5'-cyclic monophosphate; secretin receptor


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