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1 Division of Gastroenterology/Hepatology, Indiana University School of Medicine, and Richard L. Roudebush Veterans Affairs Medical Center, Indianapolis, Indiana 46202-5121; and 2 Department of Internal Medicine and Liver Center, Yale University School of Medicine, New Haven, Connecticut 06520-8019
The development of genetically altered murine animals has
generated a need for in vitro systems in the mouse. We have now characterized a novel isolated bile duct unit (IBDU) preparation from
the mouse to facilitate such studies. The mouse IBDU is isolated by
portal perfusion of collagenase, blunt dissection, further enzymatic
digestions, filtering through sized mesh, and culturing on Matrigel for
16-72 h. This mouse IBDU forms a central, enclosed lumen lined by
polarized cytokeratin-19-positive cholangiocytes with numerous
microvilli on the apical membrane. The IBDU responds to
secretory stimuli, including secretin, vasoactive intestinal peptide,
IBMX, and forskolin, resulting in expansion of the central lumen from
secretion as quantified by videomicroscopy. The secretory response to
secretin is dependent on Cl
and HCO3 in
the perfusate. These findings indicate that mouse IBDUs are intact,
polarized, functional bile duct secretory units that permit quantitative measurements of fluid secretion from mouse bile duct epithelium for the first time. This method should facilitate studies of
cholangiocyte secretion in genetically altered murine animal models.
isolated bile duct unit; biliary secretion; quantitative videomicroscopy; enzymatic isolation
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