Document Detail


Functional heterogeneity of embryonic stem cells revealed through translational amplification of an early endodermal transcript.
MedLine Citation:
PMID:  20520791     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
ES cells are defined as self-renewing, pluripotent cell lines derived from early embryos. Cultures of ES cells are also characterized by the expression of certain markers thought to represent the pluripotent state. However, despite the widespread expression of key markers such as Oct4 and the appearance of a characteristic undifferentiated morphology, functional ES cells may represent only a small fraction of the cultures grown under self-renewing conditions. Thus phenotypically "undifferentiated" cells may consist of a heterogeneous population of functionally distinct cell types. Here we use a transgenic allele designed to detect low level transcription in the primitive endoderm lineage as a tool to identify an immediate early endoderm-like ES cell state. This reporter employs a tandem array of internal ribosomal entry sites to drive translation of an enhanced Yellow Fluorescent Protein (Venus) from the transcript that normally encodes for the early endodermal marker Hex. Expression of this Venus transgene reports on single cells with low Hex transcript levels and reveals the existence of distinct populations of Oct4 positive undifferentiated ES cells. One of these cells types, characterized by both the expression of the Venus transgene and the ES cells marker SSEA-1 (V(+)S(+)), appears to represent an early step in primitive endoderm specification. We show that the fraction of cells present within this state is influenced by factors that both promote and suppress primitive endoderm differentiation, but conditions that support ES cell self-renewal prevent their progression into differentiation and support an equilibrium between this state and at least one other that resembles the Nanog positive inner cell mass of the mammalian blastocysts. Interestingly, while these subpopulations are equivalently and clonally interconvertible under self-renewing conditions, when induced to differentiate both in vivo and in vitro they exhibit different behaviours. Most strikingly when introduced back into morulae or blastocysts, the V(+)S(+) population is not effective at contributing to the epiblast and can contribute to the extra-embryonic visceral and parietal endoderm, while the V(-)S(+) population generates high contribution chimeras. Taken together our data support a model in which ES cell culture has trapped a set of interconvertible cell states reminiscent of the early stages in blastocyst differentiation that may exist only transiently in the early embryo.
Authors:
Maurice A Canham; Alexei A Sharov; Minoru S H Ko; Joshua M Brickman
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, N.I.H., Intramural; Research Support, Non-U.S. Gov't     Date:  2010-05-25
Journal Detail:
Title:  PLoS biology     Volume:  8     ISSN:  1545-7885     ISO Abbreviation:  PLoS Biol.     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-06-03     Completed Date:  2010-08-24     Revised Date:  2014-02-19    
Medline Journal Info:
Nlm Unique ID:  101183755     Medline TA:  PLoS Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e1000379     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, CD15 / genetics,  metabolism
Biological Markers / metabolism*
Blastocyst / cytology,  metabolism
Cell Differentiation* / genetics,  physiology
Cell Lineage
Cells, Cultured
Embryonic Stem Cells / cytology*,  physiology
Endoderm / cytology*,  metabolism,  physiology
Gene Expression Regulation, Developmental*
Homeodomain Proteins / genetics,  metabolism
Luminescent Proteins / genetics,  metabolism*
Mice
Morula
Octamer Transcription Factor-3 / genetics,  metabolism
Recombinant Fusion Proteins / genetics,  metabolism
Transcription Factors / genetics,  metabolism
Transgenes
Grant Support
ID/Acronym/Agency:
G0701428//Medical Research Council; G0701429//Medical Research Council; Z01AG AG000656/AG/NIA NIH HHS; Z01AG000662/AG/NIA NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, CD15; 0/Biological Markers; 0/Hhex protein, mouse; 0/Homeodomain Proteins; 0/Luminescent Proteins; 0/Octamer Transcription Factor-3; 0/Pou5f1 protein, mouse; 0/Recombinant Fusion Proteins; 0/Transcription Factors
Comments/Corrections

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