CFTR EXPRESSION AND CHLORIDE SECRETION IN POLARIZED IMMORTAL HUMAN BRONCHIAL EPITHELIAL-CELLS
CFTR EXPRESSION AND CHLORIDE SECRETION IN POLARIZED IMMORTAL HUMAN BRONCHIAL EPITHELIAL-CELLS
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DOI：
10.1165/ajrcmb.10.1.7507342
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发表时间：
1994-01-01
影响因子：
6.4
通讯作者：
GRUENERT, DC
中科院分区：
文献类型：
--
作者：
COZENS, AL;YEZZI, MJ;GRUENERT, DC
在研究人类呼吸道上皮细胞的载体离子运输、分泌和分化功能方面，一个主要的限制是缺乏合适的细胞培养系统。通过原代培养上皮细胞的转化，在这一方向上取得了进展。然而，随着连续传代的增加，这些转化子往往会失去分化的特性，特别是在危机之后。本文报道了危机后SV40大T抗原转化人支气管上皮细胞系的成功建立。该细胞系16HBE14o-，保留了分化的上皮形态和功能。细胞培养显示存在紧密连接和纤毛，单层产生跨上皮阻力，在Ussing小室中测量，并保留依赖cAMP的氯离子转运的β-肾上腺素能刺激，通过Cl-36-外流或Ussing小室中的短路电流测量。这些细胞还增加了对缓激肽或钙离子载体的氯离子转运。此外，16HBE140-细胞表达囊性纤维化跨膜传导调节因子(CFTR)mRNA和蛋白的水平，分别通过Northern和Western杂交分析很容易检测到。这些细胞为研究CFTR的调控及其在调节人呼吸道上皮细胞氯离子转运中的作用以及人类呼吸道细胞生物学的其他方面提供了有价值的资源。
A major limitation in the study of vectorial ion transport, secretion, and differentiated function in the human airway epithelium has been the lack of suitable cell culture systems. Progress in this direction has been made through the transformation of primary cultured epithelial cells. However, these transformants tend to lose differentiated properties with increasing serial passage, particularly following crisis. The successful establishment of a postcrisis SV40 large T-antigen transformed epithelial cell line derived from human bronchial epithelium is described. This cell line, 16HBE14o-, retains differentiated epithelial morphology and functions. Cell cultures show the presence of tight junctions and cilia, and monolayers generate transepithelial resistance, as measured in Ussing chambers, and retain beta-adrenergic stimulation of cAMP-dependent chloride ion transport, measured either by Cl-36- efflux or as short-circuit current in Ussing chambers. The cells also increase chloride transport in response to bradykinin or calcium ionophore. In addition, 16HBE14o- cells express levels of both the cystic fibrosis transmembrane conductance regulator (CFTR) mRNA and protein readily detectable by Northern and Western hybridization analysis, respectively. These cells provide a valuable resource for studying the modulation of CFTR and its role in regulation of chloride ion transport in human airway epithelium as well as other aspects of human airway cell biology.