Incomplete reprogramming after fusion of human multipotent stromal cells and bronchial epithelial cells.
Incomplete reprogramming after fusion of human multipotent stromal cells and bronchial epithelial cells.
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人多能基质细胞和支气管上皮细胞融合后不完全重编程。
DOI：
10.1096/fj.09-152991
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Spees,JeffreyL
中科院分区：
文献类型：
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作者：
Curril,IngridM;Koide,Masayo;Yang,CalvinH;Segal,Alan;Wellman,GeorgeC;Spees,JeffreyL
骨髓来源的祖细胞可以与多种不同组织的细胞融合，包括肺细胞，特别是在损伤后。尽管有许多关于细胞融合的报道，但很少有研究检查所产生的杂交细胞的功能。我们将人多能基质细胞（hMSCs）与正常人支气管上皮细胞（NHBEs）共培养，观察到hMSC/NHBE异核体的形成。异核体表达几种上皮细胞特有的蛋白，如角蛋白和闭塞蛋白。杂交细胞还表达了维持支气管和肺泡液平衡的2个重要离子通道的mrna和蛋白：囊性纤维化跨膜传导调节剂（CFTR）和阿米罗利敏感上皮Na+通道（ENaC）。免疫细胞化学显示，CFTR在许多杂交细胞中表达，但在其他杂交细胞中不表达或低表达。全细胞膜片钳记录显示，在氯化钡存在下，在对照NHBEs和hMSC/NHBE异核体中，格列本脲敏感电流与功能CFTR通道一致。细胞总电容测量表明，异核体的膜表面积与NHBEs相似。异核子表达α-和γ-ENaC亚基，但不表达β-ENaC亚基，表明不能形成完整的ENaC通道。此外，hMSC与表达CFTR ΔF508的永活支气管细胞融合形成的杂交细胞不会导致hMSC细胞核的重编程和野生型CFTR mRNA的表达。我们的数据表明，在成年祖细胞和体细胞融合后，重编程可能是不完整的，并可能导致细胞功能的改变。-Curril， I. M, Koide， M, Yang， C. H., Segal， A., Wellman， G. C., Spees， J. L.人多能基质细胞与支气管上皮细胞融合后不完全重编程。
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.