Increased Communication Among Nasal Epithelial Cells in Air‐Liquid Interface Culture

Abstract

Objective/Hypothesis: The retinoid acid (RA) sufficient air-liquid interface (ALI) cell culture model, but not the classical submerged single layer (SSL) cell culture model, can achieve ciliary differentiation of nasal epithelial cells. Because gap junction mediated intercellular communication (GJIC) may contribute to differentiation in numerous cell types, this study compared the extent of GJIC and the expression of Connexin 43 (Cx43) in nasal epithelial cells in both SSL and ALI cultures. Methods: Cell morphology was examined via optical and scanning electron microscope, and the number of cells with ciliary beating were counted. Lucifer Yellow dye transfer test using the scrape loading method was performed to assess the GJIC. Cx43 expression was measured with reverse-transcription polymerase chain reaction (RT-PCR) and quantitative (Q)-PCR. Results: Nasal epithelial cells in ALI culture exhibited increased numbers of ciliated cells compared with SSL culture during the 3-week culture period. On day 20, GJIC was increased in ALI culture (ALI % - SSL % = 9.6 ± 1.2%, n = 5). Accordingly, Cx43 expression was increased via RT-PCR (4.22-fold) and Q-PCR (5.3 ± 1.1-fold, n = 5) examination. Conclusions: RA sufficient ALI culture manifested more differentiated nasal epithelial cell status with ciliogenesis. Cx43, being the responsible molecule for GJIC, increased in parallel. Consequently, as in primary cultured limbal epithelial cells, Cx43 expression and extent of GJIC may serve as markers for the differentiation status of nasal epithelial cells.