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

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

Journal of allergy (2012-01-31)
Ceri E Stewart, Elizabeth E Torr, Nur H Mohd Jamili, Cynthia Bosquillon, Ian Sayers
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-β-Tubulin IV antibody produced in mouse, clone ONS.1A6, ascites fluid
Sigma-Aldrich
Anti-E-Cadherin Antibody, clone 67A4, Azide Free, clone 67A4, Chemicon®, from mouse
Sigma-Aldrich
IgG1, Kappa from murine myeloma, clone MOPC 21, ascites fluid, lyophilized powder