囊性纤维化跨膜传导调节器
Ussing室
上皮
细胞生物学
氯离子通道
离子运输机
分泌物
跨上皮电位差
细胞培养
呼吸上皮
生物
紧密连接
化学
囊性纤维化
内分泌学
生物化学
遗传学
膜
作者
Alison Cozens,M J Yezzi,Karl Kunzelmann,Takashi Ohrui,Lawrence S. Chin,Kevin H. Eng,Walter E. Finkbeiner,J. H. Widdicombe,Dieter C. Gruenert
标识
DOI:10.1165/ajrcmb.10.1.7507342
摘要
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 36Cl- 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.
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