An in vitro triple cell co-culture model with primary cells mimicking the human alveolar epithelial barrier
An in vitro triple cell co-culture model with primary cells mimicking the human alveolar epithelial barrier
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DOI：
10.1016/j.ejpb.2010.10.014
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发表时间：
2011-04-01
影响因子：
4.9
通讯作者：
Rothen-Rutishauser, Barbara M.
中科院分区：
文献类型：
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作者：
Lehmann, Andrea D.;Daum, Nicole;Rothen-Rutishauser, Barbara M.
作者最近建立了三细胞共培养模型，由 A549 或 16HBE14o- 上皮细胞、人血单核细胞来源的巨噬细胞和树突状细胞组成，这为研究异生素与这些细胞的相互作用提供了可能性。含有 16HBE140 的共培养模型模拟气道上皮屏障，而 A549 共培养模型模拟肺泡 II 型上皮屏障。目前工作的目标是建立一种新的三细胞共培养模型，该模型由从人肺活检（hAEpC）分离的原代肺泡I型样细胞组成，代表更真实的肺泡上皮屏障壁，因为I型上皮细胞覆盖了> 93％的肺泡表面。使用激光扫描显微镜和透射电子显微镜，并通过确定上皮完整性，对 A549 和 16HBE140- 的单一培养物与 hAEpC 进行形态和功能比较。使用相同的方法对三细胞共培养物进行表征。 结果表明，hAEpC 的上皮完整性（平均值 +/- SD，1180 +/- 188 Omega cm(2)）高于 A549（172 +/- 59 Omega cm(2)），但与 16lBE14o- 细胞（1469 +/- 156 Omega cm(2)）相似。与 A549 (93 +/- 14 Omega cm(2)) 和 16HBE14o- (558 +/- 267 Omega cm(2)) 的三细胞共培养模型相比，hAEpC (1113 +/- 30 Omega cm(2)) 的三细胞共培养模型显示出最高的完整性。紧密连接蛋白 zonula occlusionns-1 在 hAEpC 和 16HBE14o- 中表达更为规律，但在 A549 中则不然。 hAEpC 与两种免疫细胞（即巨噬细胞和树突状细胞）结合的上皮肺泡模型将提供一种新颖且更真实的细胞共培养系统，用于研究吸入的异生物质可能的细胞相互作用及其对人肺泡 I 型上皮壁的毒性潜力。 (C) 2010 Elsevier B.V. 保留所有权利。
A triple cell co-culture model was recently established by the authors, consisting of either A549 or 16HBE14o- epithelial cells, human blood monocyte-derived macrophages and dendritic cells, which offers the possibility to study the interaction of xenobiotics with those cells. The 16HBE140- containing co-culture model mimics the airway epithelial barrier, whereas the A549 co-cultures mimic the alveolar type II-like epithelial barrier. The goal of the present work was to establish a new triple cell co-culture model composed of primary alveolar type I-like cells isolated from human lung biopsies (hAEpC) representing a more realistic alveolar epithelial barrier wall, since type I epithelial cells cover >93% of the alveolar surface. Monocultures of A549 and 16HBE140- were morphologically and functionally compared with the hAEpC using laser scanning microscopy, as well as transmission electron microscopy, and by determining the epithelial integrity. The triple cell co-cultures were characterized using the same methods.It could be shown that the epithelial integrity of hAEpC (mean +/- SD, 1180 +/- 188 Omega cm(2)) was higher than in A549 (172 +/- 59 Omega cm(2)) but similar to 16lBE14o- cells (1469 +/- 156 Omega cm(2)). The triple cell co-culture model with hAEpC (1113 +/- 30 Omega cm(2)) showed the highest integrity compared to the ones with A549 (93 +/- 14 Omega cm(2)) and 16HBE14o- (558 +/- 267 Omega cm(2)). The tight junction protein zonula occludens-1 in hAEpC and 16HBE14o- were more regularly expressed but not in A549.The epithelial alveolar model with hAEpC combined with two immune cells (i.e. macrophages and dendritic cells) will offer a novel and more realistic cell co-culture system to study possible cell interactions of inhaled xenobiotics and their toxic potential on the human alveolar type I epithelial wall. (C) 2010 Elsevier B.V. All rights reserved.