Document Detail


ES cell extract-induced expression of pluripotent factors in somatic cells.
MedLine Citation:
PMID:  19645026     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Reprogramming of somatic cells was induced by ES cell-free extract. The system relied on the transient uptake of regulatory components from a nuclear and cytoplasmic extract derived from ES cells by the nucleus of a reversibly permeabilized NIH3T3 cell. NIH3T3 cells were permeabilized by streptolysin O (SLO). Reprogramming cell-free extracts were prepared by repeatedly freezing and thawing ES cells in liquid nitrogen. After incubation in the extract for 1 hr, permeabilized NIH3T3 cells were resealed by CaCl(2) and continually cultured for weeks to assess expression of ES cell specific markers. As we observed using FACS and fluorescence microscope, the optimal SLO concentration for permeabilizing NIH3T3 cells was 25 U. After 2 weeks of culture, the treated NIH3T3 cells began to express Nanog, c-Myc, Klf4, and 6 weeks later Oct4 was detectable. However, Sox2 was detected only after 8 weeks of culture. Differentiated somatic cells could be reprogrammed in ES extract in vitro, which provides a new approach to decreasing differentiation levels in somatic cells without disturbing the DNA sequences.
Authors:
Yan-Ning Xu; Na Guan; Zhen-Dong Wang; Zhi-Yan Shan; Jing-Ling Shen; Qing-Hua Zhang; Lian-Hong Jin; Lei Lei
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Anatomical record (Hoboken, N.J. : 2007)     Volume:  292     ISSN:  1932-8494     ISO Abbreviation:  Anat Rec (Hoboken)     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-08-05     Completed Date:  2009-11-06     Revised Date:  2009-12-16    
Medline Journal Info:
Nlm Unique ID:  101292775     Medline TA:  Anat Rec (Hoboken)     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1229-34     Citation Subset:  IM    
Copyright Information:
(c) 2009 Wiley-Liss, Inc.
Affiliation:
Department of Histology and Embryology, Harbin Medical University, Nangang District, Harbin 150081, Heilongjiang Province, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Markers / analysis
Cell Differentiation
Cell Extracts / chemistry,  pharmacology
Down-Regulation / drug effects*
Embryonic Stem Cells / chemistry*,  physiology
Fibroblasts / cytology,  drug effects,  physiology
Homeodomain Proteins / metabolism
Kruppel-Like Transcription Factors / metabolism
Mice
NIH 3T3 Cells
Nuclear Reprogramming*
Octamer Transcription Factor-3 / metabolism
Pluripotent Stem Cells / cytology,  physiology*
Proto-Oncogene Proteins c-myc / metabolism
SOXB1 Transcription Factors / metabolism
Time Factors
Chemical
Reg. No./Substance:
0/Biological Markers; 0/Cell Extracts; 0/GKLF protein; 0/Homeodomain Proteins; 0/Kruppel-Like Transcription Factors; 0/Myc protein, mouse; 0/Nanog protein, mouse; 0/Octamer Transcription Factor-3; 0/Pou5f1 protein, mouse; 0/Proto-Oncogene Proteins c-myc; 0/SOXB1 Transcription Factors; 0/Sox2 protein, mouse

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


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