Document Detail

Biphasic induction of Pdx1 in mouse and human embryonic stem cells can mimic development of pancreatic beta-cells.
MedLine Citation:
PMID:  19056911     Owner:  NLM     Status:  MEDLINE    
Embryonic stem (ES) cells represent a possible source of islet tissue for the treatment of diabetes. Achieving this goal will require a detailed understanding of how the transcription factor cascade initiated by the homeodomain transcription factor Pdx1 culminates in pancreatic beta-cell development. Here we describe a genetic approach that enables fine control of Pdx1 transcriptional activity during endoderm differentiation of mouse and human ES cell. By activating an exogenous Pdx1VP16 protein in populations of cells enriched in definitive endoderm we show a distinct lineage-dependent requirement for this transcription factor's activity. Mimicking the natural biphasic pattern of Pdx1 expression was necessary to induce an endocrine pancreas-like cell phenotype, in which 30% of the cells were beta-cell-like. Cell markers consistent with the different beta-cell differentiation stages appeared in a sequential order following the natural pattern of pancreatic development. Furthermore, in mouse ES-derived cultures the differentiated beta-like cells secreted C-peptide (insulin) in response to KCl and 3-isobutyl-1-methylxanthine, suggesting that following a natural path of development in vitro represents the best approach to generate functional pancreatic cells. Together these results reveal for the first time a significant effect of the timed expression of Pdx1 on the non-beta-cells in the developing endocrine pancreas. Collectively, we show that this method of in vitro differentiation provides a template for inducing and studying ES cell differentiation into insulin-secreting cells.
Andreia S Bernardo; Candy H-H Cho; Sharon Mason; Hilary M Docherty; Roger A Pedersen; Ludovic Vallier; Kevin Docherty
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Stem cells (Dayton, Ohio)     Volume:  27     ISSN:  1549-4918     ISO Abbreviation:  Stem Cells     Publication Date:  2009 Feb 
Date Detail:
Created Date:  2009-05-21     Completed Date:  2009-08-06     Revised Date:  2013-05-27    
Medline Journal Info:
Nlm Unique ID:  9304532     Medline TA:  Stem Cells     Country:  United States    
Other Details:
Languages:  eng     Pagination:  341-51     Citation Subset:  IM    
School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen, United Kingdom.
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MeSH Terms
Activins / pharmacology
Amyloid / genetics
Blotting, Western
Bone Morphogenetic Protein 4 / pharmacology
C-Peptide / secretion
Cell Differentiation / drug effects
Cell Line
Embryonic Stem Cells / cytology*,  drug effects,  metabolism*
Flow Cytometry
Gene Expression / drug effects
HeLa Cells
Homeodomain Proteins / genetics,  metabolism*,  physiology
Insulin / genetics
Insulin-Secreting Cells / metabolism*
Islet Amyloid Polypeptide
LIM-Homeodomain Proteins
Polymerase Chain Reaction
Tamoxifen / analogs & derivatives,  pharmacology
Trans-Activators / genetics,  metabolism*,  physiology
Transcription Factors
Grant Support
//Wellcome Trust
Reg. No./Substance:
0/Amyloid; 0/Bmp4 protein, mouse; 0/Bone Morphogenetic Protein 4; 0/C-Peptide; 0/Homeodomain Proteins; 0/Insulin; 0/Islet Amyloid Polypeptide; 0/LIM-Homeodomain Proteins; 0/Nkx6-1 protein, mouse; 0/Trans-Activators; 0/Transcription Factors; 0/activin A; 0/insulin gene enhancer binding protein Isl-1; 0/pancreatic and duodenal homeobox 1 protein; 104625-48-1/Activins; 10540-29-1/Tamoxifen; 17197F0KYM/afimoxifene

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