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

Abstract

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 x cm2 while 14C-D-mannitol flux rates (Jm) simultaneously decreased. Tight junctions were shown by diffusion potential studies to be anion selective with PC1/PNa = 1.9. Transepithelial leakiness could be induced by the phorbol ester, protein kinase C 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 Jm did not significantly fluctuate or increase. Unlike the strong inverse relationship between TER and Jm during differentiation, differentiated cell layers manifested no relationship between TER and Jm. There was also much greater variability for TER values compared to Jm. 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 (Isc) and a slight increase in TER, but no significant change in Jm. A strong temperature dependence was observed not only for Isc, 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.

Pharmacokinetics of PEGylated Gold Nanoparticles: In Vitro—In Vivo Correlation

“…Thus, in vivo results for lungs were compared with 16HBE rather than A549. Results for 16HBE cells are close to those for in vivo, thus reflecting their good barrier properties already reported in literature [ 33 ].…”

Characterisation of Aspergillus fumigatus Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy

“…Direct comparisons of Aspergillus fate in A549 and 16HBE cell lines have not been previously addressed (for review [70]), despite differential activation with lipopolysaccharide [46], Fusobacterium [71], glucocorticoids [72] and cigarette smoke [73] exposure. Overall, 16HBE cells were able to take up the A. fumigatus strains ∆pksp and ∆rodA more efficiently than A549, which may be linked with increased expression of mucus on the epithelia [74]. Additionally, improved phagosome maturation in 16HBE cells compared to A549 correlated with a 10% increment in fungal killing.…”

In Vitro Investigations on Optimizing and Nebulization of IVT-mRNA Formulations for Potential Pulmonary-Based Alpha-1-Antitrypsin Deficiency Treatment

“…The human bronchial epithelial cell line used in the current study, 16HBE14o- (16HBE), is widely used as a model for respiratory epithelial diseases and pulmonary gene therapy, due to the fact that 16HBE cells retain many of the functions and morphology of differentiated bronchial epithelial cells [ 40 ]. 16HBE cells are more static compared with other fast-dividing human bronchial epithelial cells such as BEAS-2B cells, thus being more difficult to transfect and better mimicking in vivo status [ 41 ].…”