title: Ex vivohuman pancreatic slice preparations offer a valuable model for studying pancreatic exocrine biology.
abstract: A genuine understanding of human exocrine pancreas biology and pathobiology has been hampered by a lack of suitable preparations and reliance on rodent models employing dispersed acini preparations. We have developed an organotypic slice preparation of the normal portions of human pancreas obtained from cancer resections. The preparation was assessed for physiologic and pathologic responses to the cholinergic agonist carbachol (Cch) and cholecystokinin (CCK-8), including 1) amylase secretion, 2) exocytosis, 3) intracellular Ca2+responses, 4) cytoplasmic autophagic vacuole formation, and 5) protease activation. Cch and CCK-8 both dose-dependently stimulated secretory responses from human pancreas slices similar to those previously observed in dispersed rodent acini. Confocal microscopy imaging showed that these responses were accounted for by efficient apical exocytosis at physiologic doses of both agonists and by apical blockade and redirection of exocytosis to the basolateral plasma membrane at supramaximal doses. The secretory responses and exocytotic events evoked by CCK-8 were mediated by CCK-A and not CCK-B receptors. Physiologic agonist doses evoked oscillatory Ca2+increases across the acini. Supraphysiologic doses induced formation of cytoplasmic autophagic vacuoles and activation of proteases (trypsin, chymotrypsin). Maximal atropine pretreatment that completely blocked all the Cch-evoked responses did not affect any of the CCK-8-evoked responses, indicating that rather than acting on the nerves within the pancreas slice, CCK cellular actions directly affected human acinar cells. Human pancreas slices represent excellent preparations to examine pancreatic cell biology and pathobiology and could help screen for potential treatments for human pancreatitis.
authors.affiliation: From the Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Tao
authors.initials: T
authors.lastname: Liang
authors.affiliation: From the Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Subhankar
authors.initials: S
authors.lastname: Dolai
authors.affiliation: From the Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Li
authors.initials: L
authors.lastname: Xie
authors.affiliation: Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto M5G 2N2, Ontario, Canada.
authors.forename: Erin
authors.initials: E
authors.lastname: Winter
authors.affiliation: Children's Hospital of Pittsburgh of UPMC, Rangos Research Center, Pittsburgh, Pennsylvania 15224, and.
authors.forename: Abrahim I
authors.initials: AI
authors.lastname: Orabi
authors.affiliation: From the Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Negar
authors.initials: N
authors.lastname: Karimian
authors.affiliation: From the Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Laura I
authors.initials: LI
authors.lastname: Cosen-Binker
authors.affiliation: From the Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Ya-Chi
authors.initials: YC
authors.lastname: Huang
authors.affiliation: Charles Perkins Centre, University of Sydney, Camperdown, Sydney, New South Wales, 2050, Australia.
authors.forename: Peter
authors.initials: P
authors.lastname: Thorn
authors.affiliation: Toronto General Hospital Research Institute, University Health Network, University of Toronto, Toronto M5G 2N2, Ontario, Canada.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Cattral
authors.affiliation: From the Department of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada, herbert.gaisano@utoronto.ca.
authors.forename: Herbert Y
authors.initials: HY
authors.lastname: Gaisano
mesh_terms: D000328:Adult; D000368:Aged; D005089:Exocytosis; D005260:Female; D016591:Histocytological Preparation Techniques / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D046790:Pancreas, Exocrine / Q000378:metabolism* / Q000473:pathology; D010195:Pancreatitis / Q000378:metabolism* / Q000473:pathology
keywords: CCK-8; CCK-A/B Receptor; calcium imaging; carbachol; confocal microscopy; exocytosis; human exocrine pancreas; pancreas; receptor