Document Detail

Embryonic stem cells contribute to mouse chimeras in the absence of detectable cell fusion.
MedLine Citation:
PMID:  18338954     Owner:  NLM     Status:  MEDLINE    
Embryonic stem (ES) cells are capable of differentiating into all embryonic and adult cell types following mouse chimera production. Although injection of diploid ES cells into tetraploid blastocysts suggests that tetraploid cells have a selective disadvantage in the developing embryo, tetraploid hybrid cells, formed by cell fusion between ES cells and somatic cells, have been reported to contribute to mouse chimeras. In addition, other examples of apparent stem cell plasticity have recently been shown to be the result of cell fusion. Here we investigate whether ES cells contribute to mouse chimeras through a cell fusion mechanism. Fluorescence in situ hybridization (FISH) analysis for X and Y chromosomes was performed on dissociated tissues from embryonic, neonatal, and adult wild-type, and chimeric mice to follow the ploidy distributions of cells from various tissues. FISH analysis showed that the ploidy distributions in dissociated tissues, notably the tetraploid cell number, did not differ between chimeric and wild-type tissues. To address the possibility that early cell fusion events are hidden by subsequent reductive divisions or other changes in cell ploidy, we injected Z/EG (lacZ/EGFP) ES cells into ACTB-cre blastocysts. Recombination can only occur as the result of cell fusion, and the recombined allele should persist through any subsequent changes in cell ploidy. We did not detect evidence of fusion in embryonic chimeras either by direct fluorescence microscopy for GFP or by PCR amplification of the recombined Z/EG locus on genomic DNA from ACTB-cre::Z/EG chimeric embryos. Our results argue strongly against cell fusion as a mechanism by which ES cells contribute to chimeras.
Benjamin L Kidder; Leann Oseth; Shanna Miller; Betsy Hirsch; Catherine Verfaillie; Electra Coucouvanis
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cloning and stem cells     Volume:  10     ISSN:  1536-2302     ISO Abbreviation:  Cloning Stem Cells     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-05-27     Completed Date:  2008-09-04     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  101125444     Medline TA:  Cloning Stem Cells     Country:  United States    
Other Details:
Languages:  eng     Pagination:  231-48     Citation Subset:  IM    
Department of Medicine, Stem Cell Institute, Minneapolis, Minnesota, USA.
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MeSH Terms
Blastocyst / physiology
Cell Fusion*
Embryo, Mammalian / embryology,  metabolism
Embryonic Stem Cells / physiology*
In Situ Hybridization, Fluorescence
Mice, Inbred C57BL
Mice, Transgenic
Muscle, Skeletal / cytology
Neuroepithelial Cells / cytology
Recombinases / metabolism
Recombination, Genetic
X Chromosome
Y Chromosome
Grant Support
5R01-DK058295-05/DK/NIDDK NIH HHS; 5R01-HL067932-03/HL/NHLBI NIH HHS; P30 CA077598-09/CA/NCI NIH HHS
Reg. No./Substance:

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