Multilineage murine stem cells generate complex organoids to model distal lung development and disease

Organoids derived from mouse and human stem cells have recently emerged as a powerful tool to study organ development and disease. We here established a three-dimensional (3D) murine bronchioalveolar lung organoid (BALO) model that allows clonal expansion and self-organization of FACS-sorted bronchioalveolar stem cells (BASCs) upon co-culture with lung-resident mesenchymal cells. BALOs yield a highly branched 3D structure within 21 days of culture, mimicking the cellular composition of the bronchioalveolar compartment as defined by single-cell RNA sequencing and fluorescence as well as electron microscopic phenotyping. Additionally, BALOs support engraftment and maintenance of the cellular phenotype of injected tissue-resident macrophages. We also demonstrate that BALOs recapitulate lung developmental defects after knockdown of a critical regulatory gene, and permit modeling of viral infection. We conclude that the BALO model enables reconstruction of the epithelial-mesenchymal-myeloid unit of the distal lung, thereby opening numerous new avenues to study lung development, infection, and regenerative processes in vitro.

• Vazquez-Armendariz, A. I.
• Heiner, M.
• El Agha, E.
• Salwig, I.
• Hoek, A.
• Hessler, M. C.
• Shalashova, I.
• Shrestha, A.
• Carraro, G.
• Mengel, J. P.
• Gunther, A.
• Morty, R. E.
• Vadasz, I.
• Schwemmle, M.
• Kummer, W.
• Hain, T.
• Goesmann, A.
• Bellusci, S.
• Seeger, W.
• Braun, T.
• Herold, S.