Document Detail


A novel stepwise differentiation of functional pancreatic exocrine cells from embryonic stem cells.
MedLine Citation:
PMID:  20887097     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Cultivation of functional pancreatic cells isolated from adult mammalian pancreas remains difficult. We developed a differentiation protocol that gradually induced the formation of mouse pancreatic exocrine cells from embryonic stem cells (ESCs). This process mimicked in vivo pancreatic development by directing cells through definitive endoderm (DE), gut tube endoderm, and pancreatic progenitor cells to differentiated cells that expressed pancreatic exocrine enzymes. Mouse ESCs were cultured in hanging drops to form embryoid bodies. Treatment of embryoid bodies with activin A induced the formation of DE cells that expressed marker mRNAs Goosecoid and Mixl1 and that were double-positive with Foxa2 and Sox17 proteins. Subsequent treatment of the DE cells by retinoic acid induced the formation of gut tube endoderm cells that expressed the specific marker Hnf1b. Expression of Goosecoid and Mixl1 was downregulated during this period. Fibroblast growth factor 7 (FGF7) promoted differentiation of PDX1-expressing pancreatic progenitor cells that also expressed Foxa2 mRNA, an endodermal marker, suggesting derivation from the DE cells. Exocrine cell differentiation was induced with FGF7, glucagon-like peptide-1, and nicotinamide. The differentiated cells expressed mature pancreatic exocrine cell mRNAs, such as Amylase, Elastase, and Carboxypeptidase A. Additionally, they produced pancreatic elastase, amylase, carboxypeptidase A, and chymotrypsin proteins that were identified in cytoplasmic granules by immunocytochemistry. Active amylase was released into the medium. Moreover, FGF7 was associated with differentiation of pancreatic exocrine cells. The findings reported here offer a novel and effective process to develop pancreatic exocrine cells from ESCs.
Authors:
Sakiko Shirasawa; Susumu Yoshie; Tadayuki Yokoyama; Daihachiro Tomotsune; Fengming Yue; Katsunori Sasaki
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Publication Detail:
Type:  Journal Article     Date:  2010-11-04
Journal Detail:
Title:  Stem cells and development     Volume:  20     ISSN:  1557-8534     ISO Abbreviation:  Stem Cells Dev.     Publication Date:  2011 Jun 
Date Detail:
Created Date:  2011-05-26     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101197107     Medline TA:  Stem Cells Dev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1071-8     Citation Subset:  IM    
Affiliation:
1 Laboratory for Advanced Health Sciences, Bourbon Institutes of Health , BOURBON Corporation, Kashiwazaki, Niigata, Japan .
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