Journal: Viruses

Article Title: Application of Bovine Nasal Epithelial Cells as an In Vitro Model for Studying Viral Infection in the Upper Respiratory Tract

doi: 10.3390/v17091188

Figure Lengend Snippet: Characterization of bovine nasal epithelial cell cultures. The barrier function of the bovine nasal epithelial cell (BNEC) cultures was assessed by measuring the transepithelial electrical resistance (TEER) ( A ) and by dextran diffusion assays (DDA, panel ( B )). ( A ) 24 h after seeding, BNEC cultures exceeded the confluence threshold (red dotted line). ( B ) DDA was performed using 70 kDa fluorescein isothiocyanate (FITC)-labeled dextran. No diffusion of dextran molecules was detected at 48 h post seeding. EDTA-treatment was used as a positive control. PBS-treatment was used as a negative control. ( C ) IF staining of ZO-1 indicates fully formed tight junctions. ( D ) Cytokeratin 5 (KRT5) staining confirms that the cell layer consists of epithelial basal cells. Each time point represents three biological replicates ( n = 3), each with two technical replicates. Experiments were performed twice, each time with cells from an individual donor. Results are shown as the mean ± range (minimum to maximum) of one representative experiment.

Article Snippet: DDA was performed using fluorescein isothiocyanate (FITC)-labelled 70 kilodalton (kDa) dextran molecules (Invitrogen Thermo Fisher, Waltham, MA, USA).

Techniques: Diffusion-based Assay, Labeling, Positive Control, Negative Control, Staining