Fig. 1

Sampling methods and cultivation of primary airway epithelial cells. (A) Tracheal tissue was resected in the course of size adaptations of donor lungs before transplantation. Three different sampling methods were used: conventional, forceps biopsy, and cryo biopsy. Epithelial cells were isolated and cultivated in specific media during the selection phase of basal cells. Basal cells were then frozen (400,000 cells/vial) and stored. (B) Isolated basal cells were expanded in submerged culture in T75 flasks for 8 days and seeded (40,000 cells) onto cell culture inserts. At a transepithelial electrical resistance value of 224 Ω/cm², the apical medium was removed and cells were exposed to air. Cells were then cultivated for 3 weeks in air-liquid interface and harvested after 1, 2, or 3 weeks. After 3 weeks, some cells were treated with 10 µg/mL house dust mite extract (HDM) for 6–24 h to be used for functional analyzes. Figure created with Biorender