Murine embryonic stem cell-derived pancreatic acinar cells recapitulate features of early pancreatic differentiation.
OBJECTIVE
Acinar cells constitute 90% of the pancreas epithelium, are polarized, and secrete digestive enzymes. These cells play a crucial role in pancreatitis and pancreatic cancer. However, there are limited models to study normal acinar cell differentiation in vitro. The aim of this work was to generate and characterize purified populations of pancreatic acinar cells from embryonic stem (ES) cells.


METHODS
Reporter ES cells (Ela-pur) were generated that stably expressed both beta-galactosidase and puromycin resistance genes under the control of the elastase I promoter. Directed differentiation was achieved by incubation with conditioned media of cultured fetal pancreatic rudiments and adenoviral-mediated co-expression of p48/Ptf1a and Mist1, 2 basic helix-loop-helix transcription factors crucial for normal pancreatic acinar development and differentiation.


RESULTS
Selected cells expressed multiple markers of acinar cells, including digestive enzymes and proteins of the secretory pathway, indicating activation of a coordinated differentiation program. The genes coding for digestive enzymes were not regulated as a single module, thus recapitulating what occurs during in vivo pancreatic development. The generated cells displayed transient agonist-induced Ca(2+) mobilization and showed a typical response to physiologic concentrations of secretagogues, including enzyme synthesis and secretion. Importantly, these effects did not imply the acquisition of a mixed acinar-ductal phenotype.


CONCLUSIONS
These studies allow the generation of almost pure acinar-like cells from ES cells, providing a normal cell-based model for the study of the acinar differentiation program in vitro.
Rovira
Meritxell
M
Cancer Research Program, Institut Municipal d'Investigació Mèdica, Barcelona, Spain.
Delaspre
Fabien
F
Massumi
Mohammad
M
Serra
Selma A
SA
Valverde
Miguel Angel
MA
Lloreta
Josep
J
Dufresne
Marlène
M
Payré
Bruno
B
Konieczny
Stephen F
SF
Savatier
Pierre
P
Real
Francisco X
FX
Skoudy
Anouchka
A
D000681:Amylases / Q000235:genetics; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D002217:Carbachol / Q000494:pharmacology; D043422:Carboxypeptidases A / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D018679:Cholinergic Agonists / Q000494:pharmacology; D002919:Chymotrypsinogen / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D051379:Mice; D016253:Microscopy, Immunoelectron; D046790:Pancreas, Exocrine / Q000166:cytology* / Q000196:embryology*; D010196:Pancreatic Elastase / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection
D000681:Amylases; D051792:Basic Helix-Loop-Helix Proteins; D002217:Carbachol; D043422:Carboxypeptidases A; D018679:Cholinergic Agonists; D002919:Chymotrypsinogen; D010196:Pancreatic Elastase; D014157:Transcription Factors; C491312:Bhlha15 protein, mouse; C062602:transcription factor PTF1