Mouse pancreatic acinar/ductlar tissue gives rise to epithelial cultures that are morphologically,biochemically, and functionally indistinguishable from interlobular duct cell cultures |
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Authors: | Sherwood Githens Jane A Schexnayder Randy L Moses Gerene M Denning Jeffrey J Smith Marsha L Frazier |
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Institution: | (1) Department of Biological Sciences, University of New Orleans, 70148 New Orleans, LA;(2) Department of Anatomy, L.S.U. Medical Center, 70119 New Orleans, LA;(3) Department of G.I. Oncology, University of Texas/M.D. Anderson Hospital, 77030 Houston, TX;(4) Department of Pediatrics, University of Iowa, 52242 Iowa City, IA |
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Abstract: | Summary Most of the pancreatic exocrine epithelium consists of acinar and intralobular duct (ductular) cells, with the balance consisting
of interlobular and main duct cells. Fragments of mouse acinar/ductular epithelium can be isolated by partial digestion with
collagenase and purified by Ficoll density gradient centrifugation. We investigated whether previously developed culture conditions
used for duct epithelium would result in the selective survival and proliferation of ductular cells from the acinar/ductular
fragments. The fragments were cultured on nitrocellulose filters coated with extracellular matrix. After 2 to 4 wk the filters
were covered with proliferating cells resembling parallel cultures of duct epithelium by the following criteria: protein/DNA
ratio, light and electron microscopic appearance, the presence of duct markers (carbonic anhydrase CA] activity, CA II mRNA,
the cystic fibrosis transmembrane conductance regulator), the near absence of acinar cell markers (amylase and chymotrypsin),
a similar polypeptide profile after sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the presence of spontaneous
and secretin-stimulated electrogenic ion transport. Both duct and ductular epithelia formed fluid-filled cysts in collagen
gels and both could be subcultured. We conclude that acinar/ductular tissue gives rise to ductular cells in culture by some
combination of acinar cell death and/or transdifferentiation to a ductular phenotype, accompanied by proliferation of these
cells and preexisting ductular cells. These cultures may be used to investigate the properties of this part of the pancreatic
duct system, from which most of the pancreatic juice water and electrolytes probably originates. |
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Keywords: | pancreas duct cell acinar cell culture mouse carbonic anhydrase cystic fibrosis transmembrane conductance regulator short-circuit current secretin |
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