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

EMBO J. 2020 Nov 2;39(21):e103476. doi: 10.15252/embj.2019103476. Epub 2020 Sep 28.


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.

Keywords: BALO; BASC; lung organoids; stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ataxin-1 / genetics
  • Ataxin-1 / metabolism
  • Cell Differentiation / genetics
  • Cells, Cultured
  • Endothelial Cells / cytology
  • Epithelial Cell Adhesion Molecule / genetics
  • Epithelial Cell Adhesion Molecule / metabolism
  • Epithelial Cells / cytology
  • Fibroblasts
  • Humans
  • Lung / cytology
  • Lung / growth & development*
  • Lung Diseases / pathology*
  • Mesenchymal Stem Cells
  • Mice
  • Morphogenesis / genetics
  • Morphogenesis / physiology
  • Organogenesis / physiology
  • Organoids / cytology
  • Organoids / growth & development*
  • Pulmonary Alveoli / cytology
  • Pulmonary Alveoli / growth & development
  • RNA, Messenger / metabolism
  • Regeneration / genetics
  • Regeneration / physiology
  • Stem Cells / physiology*


  • Ataxin-1
  • Atxn1 protein, mouse
  • Epithelial Cell Adhesion Molecule
  • RNA, Messenger