Immortalization of bovine pancreatic duct epithelial cells.
Marino, Lucyndia R., and Calvin U. Cotton.
The Cystic Fibrosis Center, Departments of Pediatrics and
Physiology and Biophysics, Case Western Reserve University,
Cleveland, Ohio 44106
APStracts 2:0213G, 1995.
Pancreatic duct cell lines have been isolated from a number of animal
and human tumors, but none appear to express ion transport properties
expected for differentiated pancreatic duct epithelial cells. We
sought to generate an immortalized ductal cell line from well
differentiated primary cultures of bovine pancreatic duct epithelium.
Epithelial cells from the main duct of the bovine pancreas were
isolated and immortalized by transfection with a DNA construct
encoding Simian virus Large-T antigen. A single clone(BPD1) survived
negative selection and was maintaied in culture for more than 100
passages over 2 years. The cells grow readily in culture as
monolayers and express several properties characteristic of
differentiated pancreatic ductal epithelium. The cells do not appear
to form a functional tight junction complex since the transepithelial
resistance of the monolayer cultures grown on a permeable support is
less than 10Wcm2 Northern blot analysis revealed that the cells
continue to express SV40 Large-T antigen and contain significant
levels of mRNA for proteins thought to be important in
transepithelial bicarbonate secretion [carbonic anhydrase II, Cl
-/HCO3- exchanger, Na+/H+ exchanger, and cystic fibrosis transmembrane
conductance regulator (CFTR)]. In vivo pancreatic ductal secretion is
stimulated by the peptide hormone secretin. The secretin receptor is
expressed and functionally coupled to adenylate cyclase in the
immortalized cells, since secretin caused a dose-dependent
accumulation of cAMP(20 fold increase over basal levels) with an
EC50=15 nM. Elevation of intracellular cAMP by exposure of the cells
to forskolin(10[mu]M) or secretin(0.1[mu]M) increased plasma membrane
Cl- permeability, most likely mediated by activation of CFTR. The
results of these studies demonstrate that the pancreatic duct cell
line (BPD1) retains several properties exhibited by the secretory
epithelial cells that line the pancreatic ductal tree. this cell line
should prove useful for studies of expression, function, and
regulation of pancreatic duct cell proteins.
Received 28 February 1995; accepted in final form 4 October 1995.
APS Manuscript Number G90-5.
Article publication pending Am. J. Physiol. (Gastrointest. Liver
Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 6 November 95