Figure 3
From: Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
Tight-junction formation and changes in barrier function in BBEC cultures over time. Bronchial epithelial cell cultures were grown for the indicated number of days at an ALI (relative to the establishment of the ALI) and fixed in situ on the membranes. In (A) tight-junction formation was subsequently assessed using immunofluorescence staining of the tight-junction protein ZO-1 (tight junctions - green; nuclei - blue). Representative images are shown of BBECs at −3, 12, 21 and 42 days post-ALI (see also Fig. S4). In (B) junctional complexes of ex vivo bronchial epithelial tissue and day 21 post-ALI BBEC cultures were assessed by TEM. Tight-junctions (T), adherens junctions (A) and desmosomes (D) were present along the apicolateral borders of epithelial cells in both the ex vivo tissue and BBEC cultures. In addition, membranous interdigitations (arrow) were observed between adjacent cells in the BBEC cultures. In (C) tight-junction integrity during the course of epithelial cell proliferation and differentiation was assessed by measuring the TEER of BBEC cultures until day 42 post-ALI. Nine inserts were analysed per time-point and the data represents the mean +/−standard deviation from tissue derived from three different animals (n = 27).
