title: Stage specific reprogramming of mouse embryo liver cells to a beta cell-like phenotype.
abstract: We show that cultures of mouse embryo liver generate insulin-positive cells when transduced with an adenoviral vector encoding the three genes: Pdx1, Ngn3 and MafA (Ad-PNM). Only a proportion of transduced cells become insulin-positive and the highest yield occurs in the period E14-16, declining at later stages. Insulin-positive cells do not divide further although they can persist for several weeks. RT-PCR analysis of their gene expression shows the upregulation of a whole battery of genes characteristic of beta cells including upregulation of the endogenous counterparts of the input genes. Other features, including a relatively low insulin content, the expression of genes for other pancreatic hormones, and the fact that insulin secretion is not glucose-sensitive, indicate that the insulin-positive cells remain immature. The origin of the insulin-positive cells is established both by co-immunostaining for α-fetoprotein and albumin, and by lineage tracing for Sox9, which is expressed in the ductal plate cells giving rise to biliary epithelium. This shows that the majority of insulin-positive cells arise from hepatoblasts with a minority from the ductal plate cells.
authors.affiliation: Stem Cell Institute, University of Minnesota, 2001 6th Street SE, Minneapolis, Minnesota 55455, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.forename: Ersin
authors.initials: E
authors.lastname: Akinci
authors.forename: James R
authors.initials: JR
authors.lastname: Dutton
authors.forename: Anannya
authors.initials: A
authors.lastname: Banga
authors.forename: Jonathan M W
authors.initials: JM
authors.lastname: Slack
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000418:Albumins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics*; D004622:Embryo, Mammalian; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005947:Glucose / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D051473:Maf Transcription Factors, Large / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D000509:alpha-Fetoproteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000418:Albumins; D051792:Basic Helix-Loop-Helix Proteins; D005947:Glucose; D018398:Homeodomain Proteins; D007328:Insulin; D051473:Maf Transcription Factors, Large; D009419:Nerve Tissue Proteins; D055758:SOX9 Transcription Factor; D015534:Trans-Activators; D000509:alpha-Fetoproteins; C485160:Mafa protein, mouse; C407737:Neurog3 protein, mouse; C527933:Sox9 protein, mouse; C084010:pancreatic and duodenal homeobox 1 protein
keywords: Beta cell; Hepatoblast; MafA; Ngn3; Pdx1; Sox9