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.—Curril, I. M., Koide, M., Yang, C. H., Segal, A., Wellman, G. C., Spees, J. L. Incomplete reprogramming after fusion of human multipotent stromal cells and bronchial epithelial cells. cell fusion progenitor cell CFTR ENaC Footnotes This article includes supplemental data. Please visit http://www.fasebj.org to obtain this information. Received December 24, 2009. Accepted August 5, 2010. © FASEB
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Incomplete reprogramming after fusion of human multipotent stromal cells and bronchial epithelial cells
Curril, Ingrid M.; Koide, Masayo; Yang, Calvin H.; Segal, Alan; Wellman, George C.; Spees, Jeffrey L.
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, Volume 24 (12): 4856
Fed of American Socs for Experimental Biology – Dec 1, 2010
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