Stewart Ceri E, Torr Elizabeth E, Mohd Jamili Nur H, Bosquillon Cynthia, Sayers Ian
Division of Therapeutics and Molecular Medicine, Nottingham Respiratory Biomedical Research Unit, University Hospital of Nottingham, Nottingham NG7 2UH, UK.
J Allergy (Cairo). 2012;2012:943982. doi: 10.1155/2012/943982. Epub 2012 Jan 11.
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.
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，作为用于哮喘研究的气液界面（ALI）分化模型。通过共聚焦成像和定量聚合酶链反应（qPCR）评估分化为杯状（MUC5AC+）和纤毛（β-微管蛋白IV+）细胞的能力。评估紧密连接/黏附蛋白（ZO-1、E-钙黏蛋白）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-微管蛋白IV、ZO-1和E-钙黏蛋白，并形成了TEER，然而，供体间和供体内存在很大差异。Calu-3细胞形成了更具重复性的TEER，并且具有与原代细胞相似的表型，尽管β-微管蛋白IV染色呈弥漫性。BEAS-2B细胞未分化或形成紧密连接。这些数据突出了使用原代细胞模型的挑战，以及为回答特定研究问题而仔细表征和选择系统的必要性。
