Epithelial barrier function properties of the 16HBE14o- human bronchial epithelial cell culture model

The human bronchial epithelial cell line, 16HBE14o- (16HBE), is widely used as a model for respiratory epithelial diseases and barrier function. During differentiation, transepithelial electrical resistance (TER) increased to approximately 800 Ohms × cm(2), while (14)C-d-mannitol flux rates (J(m)) simultaneously decreased. Tight junctions (TJs) were shown by diffusion potential studies to be anion-selective with P(C1)/P(Na) = 1.9. Transepithelial leakiness could be induced by the phorbol ester, protein kinase C (PKC) activator, 12-O-tetradecanoylphorbol-13-acetate (TPA), and the proinflammatory cytokine, tumor necrosis factor-α (TNF-α). Basal barrier function could not be improved by the micronutrients, zinc, or quercetin. Of methodological significance, TER was observed to be more variable and to spontaneously, significantly decrease after initial barrier formation, whereas J(m) did not significantly fluctuate or increase. Unlike the strong inverse relationship between TER and J(m) during differentiation, differentiated cell layers manifested no relationship between TER and J(m). There was also much greater variability for TER values compared with J(m). Investigating the dependence of 16HBE TER on transcellular ion conductance, inhibition of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) chloride channel with GlyH-101 produced a large decrease in short-circuit current (I(sc)) and a slight increase in TER, but no significant change in J(m). A strong temperature dependence was observed not only for I(sc), but also for TER. In summary, research utilizing 16HBE as a model in airway barrier function studies needs to be aware of the complexity of TER as a parameter of barrier function given the influence of CFTR-dependent transcellular conductance on TER.