D1.405 - Establishment of a Mouse Nasal Epithelial Cell Air–Liquid Interface Culture System and Its Application in Mucosal Barrier Research

Air–liquid interface (ALI) cultures of nasal epithelial cells are widely used to investigate nasal mucosal biology and disease mechanisms. However, the clinical availability and inflammatory status of human nasal tissues limit their broader application. To date, a standardized and reproducible ALI culture system for mouse nasal epithelial cells has not been well established, restricting mechanistic studies, particularly those involving genetically modified mouse models. 

Nasal septal mucosa was harvested from mice and enzymatically digested to isolate primary nasal epithelial cells. Cells were expanded in vitro and subsequently differentiated under ALI conditions. Epithelial–pathogen interaction models were generated by exposing ALI cultures to recombinant interleukin-13 (IL-13) and house dust mite extracts. Transepithelial electrical resistance (TEER) measurements were used to evaluate epithelial barrier integrity. Hematoxylin and eosin (H&E) staining and immunofluorescence analyses were performed to assess epithelial morphology, cellular differentiation, and the expression of barrier-associated proteins.

We established a robust and reproducible ALI culture system for mouse nasal epithelial cells, accompanied by optimized protocols for culture and downstream assays. H&E and immunofluorescence staining demonstrated the presence of differentiated ciliated epithelial cells and well-organized tight junction proteins, closely recapitulating the structural features of the nasal epithelium in vivo. Exposure to IL-13 and house dust mite extracts resulted in a significant reduction in TEER values, indicating impaired epithelial barrier function. These functional changes were accompanied by pronounced epithelial remodeling and decreased expression of mucosal barrier–related markers.

This study presents a reliable mouse nasal epithelial cell ALI culture platform that faithfully mimics in vivo nasal mucosal architecture and barrier function. The system is suitable for modeling epithelial injury and remodeling and provides a versatile tool for mechanistic studies, particularly when combined with gene-edited mouse models, thereby facilitating future research into nasal mucosal diseases.