Isolation of functional polarized bile duct units from mouse liver.
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.
Cho
W K
WK
Division of Gastroenterology/Hepatology, Indiana University School of Medicine, and Richard L. Roudebush Veterans Affairs Medical Center, Indianapolis, Indiana 46202-5121, USA. wkcho@iupui.edu
Mennone
A
A
Boyer
J L
JL
D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology; D001653:Bile Ducts, Intrahepatic / Q000166:cytology / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D002712:Chlorides / Q000494:pharmacology; D046508:Culture Techniques; D004594:Electrophysiology; D006652:Histological Techniques; D007150:Immunohistochemistry; D007633:Keratins / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008854:Microscopy, Electron; D018715:Microscopy, Video; D010477:Perfusion; D010812:Physical Stimulation; D012633:Secretin / Q000494:pharmacology
D001639:Bicarbonates; D002712:Chlorides; D012633:Secretin; D007633:Keratins