Agarose-free protocol for preparing human organotypic lung cultures to study respiratory virus infection and evaluate antivirals ex vivo

Here, we present a protocol to generate human organotypic lung cultures from surgical lung resection for the study of respiratory infections. We describe steps for tissue harvesting, biopsy punching, mechanical slicing, and culture at the air-liquid interface. This technique enables robust ex vivo infections of human lung tissues with respiratory viruses, including influenza A and Nipah. It can be used to study host-pathogen interactions, analyze innate immune responses, and evaluate antiviral candidates in physiologically relevant human lung tissue. For complete details on the use and execution of this protocol, please refer to Cezard et al. 1