Document Detail


Proteomics analysis of epithelial cells reprogrammed in cell-free extract.
MedLine Citation:
PMID:  19252170     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The functional reprogramming of a differentiated cell to a pluripotent state presents potential beneficial applications in regenerative medicine. We report here the proteomic profile of 293T epithelial cells reprogrammed to a pluripotent state using undifferentiated embryonal carcinoma (NCCIT) cellular extracts. 293T cells were reversibly permeabilized with streptolysin O, incubated in an extract of NCCIT cells or a control extract of 293T cells for 1 h, resealed with CaCl(2), and cultured. OCT4 and SOX2 gene expression were up-regulated in NCCIT extract-treated cells relative to control cells, whereas there was no alteration in DNMT3B gene expression. Thirty percent of NCCIT extract-treated cells were positive for SSEA-4, and karyotyping confirmed their 293T origin, excluding the possibility of contamination from NCCIT cells. Two-dimensional PAGE revealed approximately 400 protein spots for each cell type studied. At least 10 protein spots in the proteome of NCCIT extract-treated cells had an expression profile similar to that of NCCIT and remained unaltered in control cells. Using tandem mass spectrometry, we identified these proteins, which include 78-kDa glucose-regulated protein precursor and tropomyosin alpha-3 chain. This investigation provides the first evidence that proteins are altered in a specific manner in NCCIT extract-treated cells. This is the first report on the proteomic characterization of the nuclear reprogramming process.
Authors:
Emma Pewsey; Christine Bruce; A Stephen Georgiou; Mark Jones; Duncan Baker; Saw Yen Ow; Phillip C Wright; Christel K Freberg; Philippe Collas; Alireza Fazeli
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-02-27
Journal Detail:
Title:  Molecular & cellular proteomics : MCP     Volume:  8     ISSN:  1535-9484     ISO Abbreviation:  Mol. Cell Proteomics     Publication Date:  2009 Jun 
Date Detail:
Created Date:  2009-06-04     Completed Date:  2009-09-01     Revised Date:  2013-06-02    
Medline Journal Info:
Nlm Unique ID:  101125647     Medline TA:  Mol Cell Proteomics     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1401-12     Citation Subset:  IM    
Affiliation:
Academic Unit of Reproductive and Developmental Medicine, University of Sheffield, Level 4, The Jessop Wing, Sheffield S102SF, United Kingdom.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Cell Line
Cell-Free System
Chromatography, Liquid
DNA Primers
Electrophoresis, Gel, Two-Dimensional
Epithelial Cells / metabolism*
Flow Cytometry
Gene Expression Profiling
Humans
Karyotyping
Polymerase Chain Reaction
Proteomics*
Spectrometry, Mass, Electrospray Ionization
Tandem Mass Spectrometry
Up-Regulation
Chemical
Reg. No./Substance:
0/DNA Primers
Comments/Corrections

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


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