TRANS-EPITHELIAL TRANSPORT BY PULMONARY ALVEOLAR TYPE-II CELLS IN PRIMARY CULTURE
TRANS-EPITHELIAL TRANSPORT BY PULMONARY ALVEOLAR TYPE-II CELLS IN PRIMARY CULTURE
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DOI：
10.1073/pnas.79.19.6033
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发表时间：
1982-01-01
期刊：
影响因子：
--
通讯作者：
BERRY, LC
中科院分区：
文献类型：
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作者：
MASON, RJ;WILLIAMS, MC;BERRY, LC
体外上皮细胞的液体和电解质运输可以通过培养细胞形成圆顶的能力和单层培养物的电特性来识别。肺泡上皮细胞被认为对胎儿肺中的液体运动负有部分责任，但它们在成人肺中电解质运输中的作用尚不清楚。从成年大鼠肺中分离出 II 型肺泡细胞，并将其维持在单独的塑料培养皿上、涂有细胞外基质的塑料培养皿上以及胶原涂层的 Millipore 过滤器上。涂有细胞外基质的培养皿上形成许多大圆顶；在未涂覆的塑料培养皿上形成较小的圆顶。丁酸钠 (3 mM) 刺激圆顶形成。透射电子显微镜显示，上皮细胞已变平，但仍保留层状包涵体，并且细胞在面向培养基的顶端表面上因微绒毛而极化。在胶原蛋白涂层微孔过滤器上维持的单层膜的电特性在两个实验室进行了测试。跨上皮电位差为0.7±。 0.1 mV（24 个滤波器，7 个实验）和 1.3 .+-。 0.1 mV（13个滤光片，2个实验）心尖侧负，对应电阻为217.+-。 11 和 233 .+-。 12 欧姆/平方厘米。特布他林（10μM）产生双相反应，电位差短暂降低，然后持续增加。阿米洛利 (0.1 mM) 当添加到顶端侧时完全消除了电位差，但添加到基底侧时则不然，而 1 mM 哇巴因从基底侧更有效地抑制电位差。 II 型细胞在培养物中形成极化上皮，这些细胞在体外主动运输电解质。
Fluid and electrolyte transport by epithelial cells in vitro can be recognized by the ability of cultured cells to form domes and by the electrical properties of monolayer cultures. Pulmonary alveolar epithelial cells are thought to be partially responsible for fluid movement in the fetal lung, but their role in electrolyte transport in the adult lung is not known. Alveolar type II cells were isolated from adult rat lung and maintained on plastic culture dishes alone, on plastic culture dishes coated with an extracellular matrix and on collagen-coated Millipore filters. Numerous large domes were formed on culture dishes coated with the extracellular matrix; smaller domes were formed on uncoated plastic culture dishes. Sodium butyrate (3 mM) stimulated dome formation. Transmission electron microscopy showed that the epithelial cells had flattened but still retained lamellar inclusions and that the cells were polarized with microvilli on the apical surface facing the culture medium. The electrical properties of the monolayers maintained on collagen-coated Millipore filters were tested in 2 laboratories. The transepithelial potential differences were 0.7 .+-. 0.1 mV (24 filters, 7 experiments) and 1.3 .+-. 0.1 mV (13 filters, 2 experiments) apical side negative, and the corresponding resistances were 217 .+-. 11 and 233 .+-. 12 ohm/cm2. Terbutaline (10 .mu.M) produced a biphasic response with a transient decrease and then a sustained increase in potential difference. Amiloride (0.1 mM) completely abolished the potential difference when it was added to the apical side but not when it was added to the basal side, whereas 1 mM ouabain inhibited the potential difference more effectively from the basal side. Type II cells form a polarized epithelium in culture and these cells actively transport electrolytes in vitro.