Canus, L.; Jacolin, F.; Vasseur, V.; Cezard, A.; Ogire, E.; Aublin-Gex, A.; Bourgeais, A.; David, C.; Erny, A.; Archer, F.; Legras, A.; Sizaret, D.; Guillon, A.; Lotteau, V.; Vidalain, P.-O.; Si-Tahar, M.; Perrin-Cocon, L.; Mathieu, C.

We describe a standardized and reproducible procedure to generate human organotypic lung cultures from surgical lung resection for the study of respiratory infections. The protocol details tissue harvesting, biopsy punching, mechanical slicing, culture at the air-liquid interface. This technique enables robust ex vivo infections of human lung tissue with respiratory viruses, including Influenza A and Nipah. The described system can be used to study host-pathogen interactions, analyze innate immune responses, and evaluate antiviral candidates in physiologically relevant human lung tissue.