Document Detail


Dominance of parental genomes in embryonic stem cell/fibroblast hybrid cells depends on the ploidy of the somatic partner.
MedLine Citation:
PMID:  20512590     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Two dozen hybrid clones were produced by fusion of diploid embryonic stem (ES) cells positive for green fluorescent protein (GFP) with tetraploid fibroblasts derived from DD/c and C57BL-I(I)1RK mice. Cytogenetic analysis demonstrated that most cells from these hybrid clones contained near-hexaploid chromosome sets. Additionally, the presence of chromosomes derived from both parental cells was confirmed by polymerase chain reaction (PCR) analysis of polymorphic microsatellites. All hybrid cells were positive for GFP and demonstrated growth characteristics and fibroblast-like morphology. In addition, most hybrid cells were positive for collagen type I, fibronectin, and lamin A/C but were negative for Oct4 and Nanog proteins. Methylation status of the Oct4 and Nanog gene promoters was evaluated by bisulfite genomic sequencing analysis. The methylation sites (CpG-sites) of the Oct4 and Nanog gene promoters were highly methylated in hybrid cells, whereas the CpG-sites were unmethylated in the parental ES cells. Thus, the fibroblast genome dominated the ES genome in the diploid ES cell/tetraploid fibroblast hybrid cells. Immunofluorescent analysis of the pluripotent and fibroblast markers demonstrated that establishment of the fibroblast phenotype occurred shortly after fusion and that the fibroblast phenotype was further maintained in the hybrid cells. Fusion of karyoplasts and cytoplast derived from tetraploid fibroblasts with whole ES cells demonstrated that karyoplasts were able to establish the fibroblast phenotype of the reconstructed cells but not fibroblast cytoplasts. Thus, these data suggest that the dominance of parental genomes in hybrid cells of ES cell/somatic cell type depends on the ploidy of the somatic partner.
Authors:
Anna A Kruglova; Natalia M Matveeva; Maria M Gridina; Nariman R Battulin; Anton Karpov; Elena V Kiseleva; Ksenia N Morozova; Oleg L Serov
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-05-30
Journal Detail:
Title:  Cell and tissue research     Volume:  340     ISSN:  1432-0878     ISO Abbreviation:  Cell Tissue Res.     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-06-09     Completed Date:  2010-09-01     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0417625     Medline TA:  Cell Tissue Res     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  437-50     Citation Subset:  IM    
Affiliation:
Department of Developmental Genetics, Institute of Cytology and Genetics, Academy of Sciences of Russia, Siberian Branch, Novosibirsk, Russia.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Chromosomes, Mammalian / metabolism
Collagen Type I / genetics,  metabolism
DNA Methylation / genetics
Embryonic Stem Cells / cytology,  metabolism*,  ultrastructure
Fibroblasts / cytology,  metabolism*,  ultrastructure
Fibronectins / genetics,  metabolism
Fluorescent Antibody Technique
Gene Expression Regulation
Genome / genetics*
Homeodomain Proteins / metabolism
Hybrid Cells / cytology,  metabolism*,  ultrastructure
Lamin Type A / metabolism
Mice
Molecular Sequence Data
Octamer Transcription Factor-3 / metabolism
Ploidies*
Chemical
Reg. No./Substance:
0/Collagen Type I; 0/Fibronectins; 0/Homeodomain Proteins; 0/Lamin Type A; 0/Nanog protein, mouse; 0/Octamer Transcription Factor-3

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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