Document Detail


Incomplete reprogramming after fusion of human multipotent stromal cells and bronchial epithelial cells.
MedLine Citation:
PMID:  20724526     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Bone marrow-derived progenitor cells can fuse with cells of several different tissues, including lung, especially following injury. Despite many reports of cell fusion, few studies have examined the function of the resulting hybrid cells. We cocultured human multipotent stromal cells (hMSCs) and normal human bronchial epithelial cells (NHBEs) and observed the formation of hMSC/NHBE heterokaryons. The heterokaryons expressed several proteins characteristic of epithelial cells, such as keratin and occludin. Hybrid cells also expressed the mRNAs and proteins for 2 important ion channels that maintain bronchial and alveolar fluid balance: the cystic fibrosis transmembrane conductance regulator (CFTR) and the amiloride-sensitive epithelial Na(+) channel (ENaC). By immunocytochemistry, CFTR was expressed in many hybrid cells but was absent or low in others. Whole-cell patch-clamp recordings demonstrated a glibenclamide-sensitive current in the presence of barium chloride, consistent with functional CFTR channels, in control NHBEs and hMSC/NHBE heterokaryons. Total cell capacitance measurements showed that the membrane surface area of heterokaryons was similar to that of NHBEs. Heterokaryons expressed the α- and γ-ENaC subunits but did not express the β-ENaC subunit, indicating the inability to form a complete ENaC channel. In addition, hybrid cells formed by the fusion of hMSCs with immortalized bronchial cells that expressed CFTR ΔF508 did not lead to reprogramming of the hMSC nucleus and expression of wild-type CFTR mRNA. Our data show that reprogramming can be incomplete following fusion of adult progenitor cells and somatic cells and may lead to altered cell function.
Authors:
Ingrid M Curril; Masayo Koide; Calvin H Yang; Alan Segal; George C Wellman; Jeffrey L Spees
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-08-19
Journal Detail:
Title:  FASEB journal : official publication of the Federation of American Societies for Experimental Biology     Volume:  24     ISSN:  1530-6860     ISO Abbreviation:  FASEB J.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-12-02     Completed Date:  2011-01-04     Revised Date:  2013-05-28    
Medline Journal Info:
Nlm Unique ID:  8804484     Medline TA:  FASEB J     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4856-64     Citation Subset:  IM    
Affiliation:
Cell and Molecular Biology Program and Vermont Lung Center, Burlington, Vermont, USA.
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MeSH Terms
Descriptor/Qualifier:
Bronchi / cytology
Cell Fusion
Cell Membrane / metabolism
Cells, Cultured
Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
Electrophysiology
Epithelial Cells / cytology*,  metabolism*
Flow Cytometry
Humans
Immunohistochemistry
Multipotent Stem Cells / cytology*,  metabolism*
Nuclear Reprogramming / genetics,  physiology*
Reverse Transcriptase Polymerase Chain Reaction
Stromal Cells / cytology*,  metabolism*
Grant Support
ID/Acronym/Agency:
P20 RR-155557/RR/NCRR NIH HHS; R01-HL-085210/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
126880-72-6/Cystic Fibrosis Transmembrane Conductance Regulator
Comments/Corrections

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


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