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1 Department of Internal Medicine, Scott and White Hospital, Temple, TX, USA; Division of Medical Physiology, The Texas A&M University System Health Science Center College of Medicine, Temple, TX, USA; Central Texas Veterans Health Care System, Temple, TX, USA
2 Division of Research and Education, The Texas A&M University System Health Science Center College of Medicine, Temple, TX, USA
3 Department of Gastroenterology, University of Ancona, Ancona, Italy
4 Department of Internal Medicine, Scott and White Hospital, Temple, TX, USA; Department of Medical Biochemistry and Genetics, Scott and White Hospital, Temple, Tx, USA
* To whom correspondence should be addressed. E-mail: gene.lesage{at}uth.tmc.edu.
We sought to develop a cholangiocyte cell culture system that has preservation of receptors, transporters and channels that are involved in secretin-induced secretion. METHODS: Intrahepatic bile duct fragments, obtained by enzyme perfusion of normal rat liver, were seeded on collagen and maintained in culture up to 18 weeks. 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 (CFTR) and Cl-/HCO3- exchanger and 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. RESULTS: Cultured cholangiocytes were all gamma-GT and CK-19 positive. The cells expressed SR and Cl-/HCO3- exchanger and secretin-stimulated cAMP synthesis, Cl-/HCO3- 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. SUMMARY/CONCLUSIONS: We have developed a novel cholangiocyte line that has persistent HCO3-, Cl-, and fluid transport functions. This cell system should be useful to investigators who study cholangiocyte secretion.
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