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首页> 外文期刊>Infection and immunity >A549 Lung Epithelial Cells Grown as Three-Dimensional Aggregates: Alternative Tissue Culture Model for Pseudomonas aeruginosa Pathogenesis
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A549 Lung Epithelial Cells Grown as Three-Dimensional Aggregates: Alternative Tissue Culture Model for Pseudomonas aeruginosa Pathogenesis

机译：三维聚集体生长的A549肺上皮细胞：铜绿假单胞菌发病机理的替代组织培养模型

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A three-dimensional (3-D) lung aggregate model was developed from A549 human lung epithelial cells by using a rotating-wall vessel bioreactor to study the interactions between Pseudomonas aeruginosa and lung epithelial cells. The suitability of the 3-D aggregates as an infection model was examined by immunohistochemistry, adherence and invasion assays, scanning electron microscopy, and cytokine and mucoglycoprotein production. Immunohistochemical characterization of the 3-D A549 aggregates showed increased expression of epithelial cell-specific markers and decreased expression of cancer-specific markers compared to their monolayer counterparts. Immunohistochemistry of junctional markers on A549 3-D cells revealed that these cells formed tight junctions and polarity, in contrast to the cells grown as monolayers. Additionally, the 3-D aggregates stained positively for the production of mucoglycoprotein while the monolayers showed no indication of staining. Moreover, mucin-specific antibodies to MUC1 and MUC5A bound with greater affinity to 3-D aggregates than to the monolayers. P. aeruginosa attached to and penetrated A549 monolayers significantly more than the same cells grown as 3-D aggregates. Scanning electron microscopy of A549 cells grown as monolayers and 3-D aggregates infected with P. aeruginosa showed that monolayers detached from the surface of the culture plate postinfection, in contrast to the 3-D aggregates, which remained attached to the microcarrier beads. In response to infection, proinflammatory cytokine levels were elevated for the 3-D A549 aggregates compared to monolayer controls. These findings suggest that A549 lung cells grown as 3-D aggregates may represent a more physiologically relevant model to examine the interactions between P. aeruginosa and the lung epithelium during infection.
机译：利用旋转壁血管生物反应器，从A549人肺上皮细胞建立了三维（3-D）肺聚集模型，以研究铜绿假单胞菌与肺上皮细胞之间的相互作用。通过免疫组织化学，粘附和侵袭试验，扫描电子显微镜以及细胞因子和粘糖蛋白的产生来检验3-D聚集体作为感染模型的适用性。 3-D A549聚集体的免疫组织化学表征显示，与单层类似物相比，上皮细胞特异性标记物表达增加，而癌症特异性标记物表达减少。与单层生长的细胞相比，A549 3-D细胞上的连接标记的免疫组织化学显示这些细胞形成紧密的连接和极性。另外，3-D聚集体对粘糖蛋白的产生染色呈阳性，而单层膜未显示染色迹象。而且，针对MUC1和MUC5A的粘蛋白特异性抗体与3-D聚集体的结合亲和力比与单层结合的亲和力更大。 P。铜绿假单胞菌附着并穿透A549单层的能力明显高于以3-D聚集体生长的相同细胞。扫描电镜观察A549细胞生长成单层细胞并感染了 P的3-D聚集体。铜绿假单胞菌显示感染后单层从培养板表面脱落，而3-D聚集体则仍附着在微载体珠上。响应感染，与单层对照相比，3-D A549聚集体的促炎细胞因子水平升高。这些发现表明，生长成3-D聚集体的A549肺细胞可能代表了一种更具生理相关性的模型，以检验 P之间的相互作用。铜绿假单胞菌和肺上皮感染。

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