Document Detail


Comparison of reprogramming efficiency between transduction of reprogramming factors, cell-cell fusion, and cytoplast fusion.
MedLine Citation:
PMID:  20572011     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Reprogramming human somatic cells into pluripotent cells opens up new possibilities for transplantation therapy, the study of disease, and drug screening. In addition to somatic cell nuclear transfer, several approaches to reprogramming human cells have been reported: transduction of defined transcription factors to generate induced pluripotent stem cell (iPSC), human embryonic stem cell (hESC)-somatic cell fusion, and hESC cytoplast-somatic cell fusion or exposure to extracts of hESC. Here, we optimized techniques for hESC-human fibroblast fusion and enucleation and cytoplast fusion, and then compared the reprogramming efficiency between iPSC generation, cell-fusion and cytoplast-fusion. When compared with iPSC, hESC-fusion provided much faster and efficient reprogramming of somatic cells. The reprogramming required more than 4 weeks and the efficiency was less than 0.001% in iPSC generation, and it was less than 10 days and more than 0.005% in hESC-fusion. In addition, fusion yielded almost no partially reprogrammed cell colonies. However, the fused cells were tetraploid or aneuploid. hESC cytoplast fusion could initiate reprogramming but was never able to complete reprogramming. These data indicate that in cell fusion, as in nuclear transfer, reprogramming through direct introduction of a somatic nucleus into the environment of a pluripotent cell provides relatively efficient reprogramming. The findings also suggest that the nucleus of the host pluripotent cell may contain components that accelerate the reprogramming process.
Authors:
Kouichi Hasegawa; Peilin Zhang; Zong Wei; Jordan E Pomeroy; Wange Lu; Martin F Pera
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Stem cells (Dayton, Ohio)     Volume:  28     ISSN:  1549-4918     ISO Abbreviation:  Stem Cells     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-08-25     Completed Date:  2011-01-13     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9304532     Medline TA:  Stem Cells     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1338-48     Citation Subset:  IM    
Affiliation:
Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research at USC, University of Southern California, Los Angeles, California 90033, USA. kouichi@med.usc.edu
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MeSH Terms
Descriptor/Qualifier:
Cell Differentiation / genetics,  physiology*
Cell Fusion / methods*
Cell Line
Embryonic Stem Cells / cytology*,  metabolism*
Genetic Vectors / genetics
Humans
Induced Pluripotent Stem Cells
Kruppel-Like Transcription Factors / genetics
Lentivirus / genetics
Nuclear Reprogramming / genetics,  physiology*
Octamer Transcription Factor-3 / genetics
Proto-Oncogene Proteins c-myc / genetics
SOXB1 Transcription Factors / genetics
Transduction, Genetic / methods*
Chemical
Reg. No./Substance:
0/GKLF protein; 0/Kruppel-Like Transcription Factors; 0/MYC protein, human; 0/Octamer Transcription Factor-3; 0/POU5F1 protein, human; 0/Proto-Oncogene Proteins c-myc; 0/SOX2 protein, human; 0/SOXB1 Transcription Factors

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


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