P036 A comparative study of cell culture inserts for in vitro modeling of the cystic fibrosis airway epithelium

Objectives: To generate physiologically relevant models to study the human airways, researchers are relying on cell cultures grown as organoids in hydrogel or Air-liquid interface (ALI) transwells. A consequence of coronavirus (COVID-19) pandemic and current war Ukraine, have been a global shortage lab consumables including Corning Transwells, typically used for airway ALI cultures. One way overcome this supply issue, is use alternative culture inserts. In study, we compared available commercial inserts, ensure exchangeability reproducibility across brands. Methods: Airway epithelial basal cells were seeded inserts differentiation was monitored over 4 week period. We an immortalized line primary from CF non-CF airways. After weeks culture, measured layer thickness, integrity, cellular composition. The additionally tested infection models, measuring bacterial penetration, cytotoxicity, cytokine secretion upon 14 hours co-culture with bacteria specifically prevalent Results: found significant morphological differences different included variation both thickness status. Several supported no only limited cells. Upon infection, detected localization cytotoxicity levels off secretion. Conclusions: Comparing systems essential experiments before changing components. Choice system may severely influence disease modeling host-microbe interaction studies. did not support cells, limits our ability model airways vitro, severe negative impact future research.