title: Viral-mediated coexpression of Pdx1 and p48 regulates exocrine pancreatic differentiation in mouse ES cells.
abstract: Embryonic stem cells (ES) can spontaneously activate a pancreatic differentiation program in vitro, although with low efficiency. The aim was to improve such process by using viral mediated gene transduction. In this study, we have examined the suitability of using viral vectors to express key transcriptional factors involved in pancreatic development. ES cell lines that constitutively express Pdx1, a homeodomain protein involved in both exocrine and endocrine pancreatic development and differentiation, were established using a lentiviral vector. These cells were additionally infected with an adenovirus expressing p48, a bHLH factor that is also crucial for pancreatic development and acinar differentiation. Quantitative RT-PCR analysis demonstrated an increase in the expression of exocrine genes, including those coding for both digestive enzymes and transcription factors. Immunocytochemical staining also revealed an increase in the number of amylase-expressing cell clusters. However, other important genes involved in acinar cell maturation (i.e., Mist1) were not modulated under these conditions, suggesting that the cells display features of immature exocrine cells or because of an uncoupled gene expression of the exocrine differentiation program. Importantly, this effect was selective for the acinar lineage as the expression of a large set of endocrine markers remained unchanged. Therefore, combined expression of key genes involved in pancreatic development may be a promising approach to generate mature pancreatic exocrine cells.
authors: Cell and Molecular Biology Unit, Institut Municipal d'Investigació Mèdica (IMIM), Dr Aiguader 88, Barcelona, Spain.
authors: Meritxell
authors: M
authors: Rovira
authors: Judit
authors: J
authors: Jané-Valbuena
authors: Mélanie
authors: M
authors: Marchand
authors: Pierre
authors: P
authors: Savatier
authors: Francisco X
authors: FX
authors: Real
authors: Anouchka
authors: A
authors: Skoudy
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D018398:Homeodomain Proteins / Q000096:biosynthesis* / Q000235:genetics; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D046790:Pancreas, Exocrine / Q000166:cytology* / Q000378:metabolism; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D018398:Homeodomain Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C084010:pancreatic and duodenal homeobox 1 protein; C062602:transcription factor PTF1