English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Induction and selection of Sox17-expressing endoderm cells generated from murine embryonic stem cells

Schroeder, I. S., Sulzbacher, S., Nolden, T., Fuchs, J., Czarnota, J., Meisterfeld, R., et al. (2012). Induction and selection of Sox17-expressing endoderm cells generated from murine embryonic stem cells. Cells Tissues Organs, 195(6), 507-523. doi:10.1159/000329864.

Item is

Files

show Files
hide Files
:
Schroeder.pdf (Publisher version), 2MB
Name:
Schroeder.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
© 2011 S. Karger AG
License:
-

Locators

show

Creators

show
hide
 Creators:
Schroeder, I. S., Author
Sulzbacher, S., Author
Nolden, T., Author
Fuchs, J., Author
Czarnota, J., Author
Meisterfeld, R., Author
Himmelbauer, H.1, 2, Author              
Wobus, A. M., Author
Affiliations:
1Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, Berlin, Germany, ou_1433550              
2Centre for Genomic Regulation, Pompeu Fabra University, Barcelona, Spain, ou_persistent22              

Content

show
hide
Free keywords: Activins/pharmacology Animals Biological Markers/metabolism Cell Differentiation/drug effects/genetics Cell Lineage/drug effects/genetics Cell Separation/*methods Embryoid Bodies/cytology/drug effects/metabolism Embryonic Stem Cells/*cytology/drug effects/*metabolism Endoderm/*cytology/drug effects/*metabolism Epithelium/drug effects/embryology/metabolism Flow Cytometry Fluorescent Antibody Technique Gene Expression Regulation, Developmental/drug effects HMGB Proteins/*metabolism Luminescent Proteins/metabolism Mice Octamer Transcription Factor-3/genetics/metabolism Pluripotent Stem Cells/cytology/drug effects/metabolism RNA, Messenger/genetics/metabolism SOXF Transcription Factors/*metabolism Time Factors
 Abstract: Embryonic stem (ES) cells offer a valuable source for generating insulin-producing cells. However, current differentiation protocols often result in heterogeneous cell populations of various developmental stages. Here we show the activin A-induced differentiation of mouse ES cells carrying a homologous dsRed-IRES-puromycin knock-in within the Sox17 locus into the endoderm lineage. Sox17-expressing cells were selected by fluorescence-assisted cell sorting (FACS) and characterized at the transcript and protein level. Treatment of ES cells with high concentrations of activin A for 10 days resulted in up to 19% Sox17-positive cells selected by FACS. Isolated Sox17-positive cells were characterized by defini- tive endoderm-specific Sox17/Cxcr4/Foxa2 transcripts, but lacked pluripotency-associated Oct4 mRNA and protein. The Sox17-expressing cells showed downregulation of extraembryonic endoderm (Sox7, Afp, Sdf1)-, mesoderm (Foxf1, Meox1)- and ectoderm (Pax6, NeuroD6)-specific transcripts. The presence of Hnf4alpha, Hes1 and Pdx1 mRNA demonstrated the expression of primitive gut/foregut cell-specific markers. Ngn3, Nkx6.1 and Nkx2.2 transcripts in Sox17-positive cells were determined as properties of pancreatic endocrine progenitors. Immunocytochemistry of activin A-induced Sox17-positive embryoid bodies revealed coexpression of Cxcr4 and Foxa2. Moreover, the histochemical demonstration of E-cadherin-, Cxcr4-, Sox9-, Hnf1beta- and Ngn3-positive epithelial-like structures underlined the potential of Sox17-positive cells to further differentiate into the pancreatic lineage. By reducing the heterogeneity of the ES cell progeny, Sox17-expressing cells are a suitable model to evaluate the effects of growth and differentiation factors and of culture conditions to delineate the differentiation process for the generation of pancreatic cells in vitro.

Details

show
hide
Language(s): eng - English
 Dates: 2012
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1159/000329864
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Cells Tissues Organs
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Basel : Karger
Pages: - Volume / Issue: 195 (6) Sequence Number: - Start / End Page: 507 - 523 Identifier: ISSN: 1422-6405
CoNE: https://pure.mpg.de/cone/journals/resource/954926935676