Evaluation of differentiated human bronchial epithelial cell culture systems for asthma research.

Journal of allergy Pub Date : 2012-01-01 Epub Date: 2012-01-11 DOI:10.1155/2012/943982
Ceri E Stewart, Elizabeth E Torr, Nur H Mohd Jamili, Cynthia Bosquillon, Ian Sayers
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引用次数: 177

Abstract

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.

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分化人支气管上皮细胞培养系统在哮喘研究中的评价。
本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作为气液界面(ALI)分化模型在哮喘研究中的应用。通过共聚焦成像和qPCR评估其向杯状(MUC5AC+)和纤毛状(β-微管蛋白IV+)细胞分化的能力。评估紧密连接/粘附蛋白(ZO-1, E-Cadherin)的表达和上皮电阻抗(TEER)的发展。原代细胞显示MUC5AC、β-微管蛋白IV、ZO-1和E-Cadherin的局部定位,并发生TEER,但在供体间和供体内存在很大程度的差异。Calu-3细胞虽然具有弥漫性β-微管蛋白IV染色,但其TEER的可重复性更高,表型与原代细胞相似。BEAS-2B细胞未分化,也未形成紧密连接。这些数据突出了使用原代细胞模型的挑战,以及需要仔细表征和选择系统来回答具体的研究问题。
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