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首页> 外文期刊>American Journal of Physiology >Fluid transport across cultured rat alveolar epithelial cells: a novel in vitro system.
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Fluid transport across cultured rat alveolar epithelial cells: a novel in vitro system.

机译：跨培养的大鼠肺泡上皮细胞的液体运输：一种新型的体外系统。

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Previous studies have used fluid-instilled lungs to measure net alveolar fluid transport in intact animal and human lungs. However, intact lung studies have two limitations: the contribution of different distal lung epithelial cells cannot be studied separately, and the surface area for fluid absorption can only be approximated. Therefore, we developed a method to measure net vectorial fluid transport in cultured rat alveolar type II cells using an air-liquid interface. The cells were seeded on 0.4-microm microporous inserts in a Transwell system. At 96 h, the transmembrane electrical resistance reached a peak level (1,530 +/- 115 Omega.cm(2)) with morphological evidence of tight junctions. We measured net fluid transport by placing 150 microl of culture medium containing 0.5 microCi of (131)I-albumin on the apical side of the polarized cells. Protein permeability across the cell monolayer, as measured by labeled albumin, was 1.17 +/- 0.34% over 24 h. The change in concentration of (131)I-albumin in the apical fluid was used to determine the net fluid transported across the monolayer over 12 and 24 h. The net basal fluid transport was 0.84 microl.cm(-2).h(-1). cAMP stimulation with forskolin and IBMX increased fluid transport by 96%. Amiloride inhibited both the basal and stimulated fluid transport. Ouabain inhibited basal fluid transport by 93%. The cultured cells retained alveolar type II-like features based on morphologic studies, including ultrastructural imaging. In conclusion, this novel in vitro system can be used to measure net vectorial fluid transport across cultured, polarized alveolar epithelial cells.
机译：先前的研究已使用滴注肺部来测量完整动物肺和人肺中的净肺泡液转运。但是，完整的肺部研究有两个局限性：不能单独研究不同的远端肺上皮细胞的贡献，并且只能近似吸收液体的表面积。因此，我们开发了一种使用气液界面测量培养的大鼠II型肺泡细胞中的矢量流的方法。将细胞接种在Transwell系统中的0.4微米微孔插入物上。在96 h时，跨膜电阻达到峰值水平（1,530 +/- 115 Omega.cm（2）），具有紧密连接的形态学证据。我们通过在极化细胞的顶侧放置150微升含有0.5微居里的（131）I-白蛋白的培养基来测量净流体输送。通过标记的白蛋白测量，在整个24小时内，穿过细胞单层的蛋白质渗透率为1.17 +/- 0.34％。顶液中（131）I-白蛋白浓度的变化用于确定在12和24小时内穿过单层转运的净液。净基础流体输送为0.84 microl.cm（-2）.h（-1）。用福司可林和IBMX刺激cAMP可使液体转运增加96％。阿米洛利既抑制基础运输又刺激液体运输。瓦巴因抑制了基础液的运输93％。根据形态学研究，包括超微结构成像，培养的细胞保留了II型肺泡特征。总之，这种新颖的体外系统可用于测量跨培养的极化肺泡上皮细胞的净矢量流体运输。

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