Document Detail

Reprogramming factors involved in hybrids and cybrids of human embryonic stem cells fused with hepatocytes.
MedLine Citation:
PMID:  20936904     Owner:  NLM     Status:  In-Process    
Embryonic stem cells (ESCs) have the potential to reprogram somatic cells into ESC-like cells through cell fusion. In the present study, the potential of human (h)ESC cytoplasts and karyoplasts to reprogram human hepatocytes was evaluated. Green fluorescent protein (GFP) transfected hESCs (ENVY cells) were fused with SNARF-1 (CellTracker)-labeled human hepatocytes using polyethylene glycol (PEG) and fluorescence-activated cell sorting (FACS) to produce hESC-hepatocyte hybrids. Immunocytochemical analysis of ESC markers showed that the hybrids expressed OCT4, TRA-1-60, TRA-1-81, SSEA-4, and GCTM-2. However, SSEA-1, which is typically low or absent on hESCs, was detected on hESC–hepatocyte hybrids. Moreover, reverse transcriptase polymerase chain reaction (RT-PCR) showed that alpha-fetoprotein, which is highly expressed in hepatocytes, was erased in the hybrids. These results indicated that hESCs have the potential to reprogram hepatocyte phenotype to a relatively undifferentiated state, but such hybrid cells are not identical to hESCs. Although hESC–hepatocyte hybrids were aneuploid, they were able to differentiate into embryoid bodies and some types of somatic cells. Furthermore, cybrids of enucleated hESCs and hepatocytes were produced by cell fusion, but the cybrids were unable to self-renew in the same way as hESCs. Presumably, the reprogramming factors are associated with the karyoplast and not the cytoplast of hESCs.
Jitong Guo; R Tayfur Tecirlioglu; Linh Nguyen; Karen Koh; Graham Jenkin; Alan Trounson
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cellular reprogramming     Volume:  12     ISSN:  2152-4998     ISO Abbreviation:  Cell Reprogram     Publication Date:  2010 Oct 
Date Detail:
Created Date:  2010-10-12     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101528176     Medline TA:  Cell Reprogram     Country:  United States    
Other Details:
Languages:  eng     Pagination:  529-41     Citation Subset:  IM    
Monash University, Melbourne, Australia.
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