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Evaluation of Differentiated Human Bronchial Epithelial Cell Culture Systems for Asthma Research

机译:差异化人支气管上皮细胞培养系统的哮喘研究评价

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摘要

The aim of the current study was to evaluate primary (human bronchial epithelial cells, HBEC) and non-primary (Calu-3, BEAS-2B, BEAS-2B R1) bronchial epithelial cell culture systems as air-liquid interface- (ALI-) differentiated models for asthma research. Ability to differentiate into goblet (MUC5AC+) and ciliated (β-Tubulin IV+) cells was evaluated by confocal imaging and qPCR. Expression of tight junction/adhesion proteins (ZO-1, E-Cadherin) and development of transepithelial electrical resistance (TEER) were assessed. Primary cells showed localised MUC5AC, β-Tubulin IV, ZO-1, and E-Cadherin and developed TEER with, however, a large degree of inter- and intradonor variation. Calu-3 cells developed a more reproducible TEER and a phenotype similar to primary cells although with diffuse β-Tubulin IV staining. BEAS-2B cells did not differentiate or develop tight junctions. These data highlight the challenges in working with primary cell models and the need for careful characterisation and selection of systems to answer specific research questions.
机译:当前研究的目的是评估原代(人支气管上皮细胞,HBEC)和非原代(Calu-3,BEAS-2B,BEAS-2B R1)支气管上皮细胞培养系统的气液界面-(ALI- )用于哮喘研究的差异化模型。通过共聚焦成像和qPCR评估分化为杯状(MUC5AC +)和纤毛(β-TubulinIV +)细胞的能力。评估了紧密连接/粘附蛋白(ZO-1,E-钙黏着蛋白)的表达和跨上皮电阻(TEER)的发展。原代细胞显示出局部的MUC5AC,β-TubulinIV,ZO-1和E-钙黏着蛋白,并发育出TEER,但供体间和供体内差异很大。尽管具有弥漫性β-微管蛋白IV染色,但Calu-3细胞的TEER和表型与原代细胞相似,可再现性更高。 BEAS-2B细胞未分化或形成紧密连接。这些数据凸显了使用原代细胞模型所面临的挑战,以及需要仔细表征和选择系统以回答特定研究问题的挑战。

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