Electrical properties of monolayers cultured from cells of human tracheal mucosa.
Electrical properties of monolayers cultured from cells of human tracheal mucosa.
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从人气管粘膜细胞培养的单层细胞的电特性。
DOI：
10.1152/jappl.1985.58.5.1729
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Tuet,IK
中科院分区：
文献类型：
--
作者：
Widdicombe,JH;Coleman,DL;Finkbeiner,WE;Tuet,IK
采用胶原酶消化法从人气管黏膜中分离得到分散的细胞。每条气管可获得1.5×10(8)个细胞，台盼蓝拒染法显示细胞存活率高达95%。当在培养中生长时，细胞在大约4天后形成单层。单分子膜的电子显微镜显示出一种极化结构。由典型的紧密连接将顶膜与相对不特化的基侧膜分开，顶膜含有微绒毛和明显的糖萼。生长在核孔过滤器上的单分子膜显示出44-1,800 omega的电阻。Cm2和跨上皮电位差为0.1-7.6 mV。异丙肾上腺素、前列腺素E_2和F_2α、缓激肽可增加短路电流(ISC)。当环状利尿剂布美他尼加入到基底外侧(浆膜)侧时，可降低ISC，但对单层的顶端(粘膜)侧无影响。速尿和MK-196对ISC也有抑制作用。阿米洛利对ISC有抑制作用。阿米洛利和哇巴因对ISC无影响。Serosal哇巴因在大约15分钟后使ISC降至零。
Dispersed isolated cells were obtained from human tracheal mucosa by digestion with collagenase. Up to 1.5 X 10(8) cells were obtained per trachea and showed up to 95% viability, as judged by trypan blue exclusion. When grown in culture, the cells formed monolayers after approximately 4 days. Electron microscopy of the monolayers revealed a polarized structure. An apical membrane, containing microvilli and a pronounced glycocalyx, was separated from a relatively unspecialized basolateral membrane by typical tight junctions. Monolayers grown on nucleopore filters showed resistances of 44–1,800 omega. cm2 and transepithelial potential differences of 0.1–7.6 mV. Short-circuit current (Isc) was increased by isoproterenol, prostaglandins E2 and F2 alpha, and bradykinin. The loop diuretic, bumetanide, reduced Isc when added to the basolateral (serosal) side but had no effect from the apical (mucosal) side of the monolayers. Furosemide and MK-196 also inhibited Isc. Mucosal amiloride inhibited Isc. Serosal amiloride or mucosal ouabain had no effect on Isc. Serosal ouabain brought Isc to zero after approximately 15 min.