Abstract: Compositions and methods are provided for the culture of mammalian distal lung organoids that robustly preserve and propagate distal lung alveoli and bronchioles en bloc with infiltrating endogenous immune cells and mesenchyme as a cohesive unit without artificial reconstitution, allowing analysis of the dynamics of tissue-level inflammation upon respiratory infection.

Abstract: Abstract: We describe a robust human adult distal lung organoid method with a procedure for everting organoids to essentially turn them inside out. This then relocates the apical ACE2-expressing surfaces of cells to the organoid exterior, where they can then be easily infected by SARS-CoV-2 added to the tissue culture medium. Further, this method can be used for infection of any distal lung pathogen that infects apically. Alternatively, if a pathogen interacts basolaterally then eversion is not necessary, and the human adult distal lung organoids can be infected as is.