Recapitulation of lung development from human pluripotent stem cells (hPSCs) in three dimensions (3D) would allow deeper insight into human development, as well as the development of innovative strategies for disease modelling, drug discovery and regenerative medicine1. We report here the generation from hPSCs of lung bud organoids (LBOs) that contain mesoderm and pulmonary endoderm and develop into branching airway and early alveolar structures after xenotransplantation and in Matrigel 3D culture. Expression analysis and structural features indicated that the branching structures reached the second trimester of human gestation. Infection in vitro with respiratory syncytial virus, which causes small airway obstruction and bronchiolitis in infants2, led to swelling, detachment and shedding of infected cells into the organoid lumens, similar to what has been observed in human lungs3. Introduction of mutation in HPS1, which causes an early-onset form of intractable pulmonary fibrosis4,5, led to accumulation of extracellular matrix and mesenchymal cells, suggesting the potential use of this model to recapitulate fibrotic lung disease in vitro. LBOs therefore recapitulate lung development and may provide a useful tool to model lung disease.

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  1. 1.

    & Organogenesis in a dish: modeling development and disease using organoid technologies. Science 345, 1247125 (2014).

  2. 2.

    , & Respiratory syncytial virus: virology, reverse genetics, and pathogenesis of disease. Curr. Top. Microbiol. Immunol. 372, 3–38 (2013).

  3. 3.

    , , , & The histopathology of fatal untreated human respiratory syncytial virus infection. Mod. Pathol. 20, 108–119 (2007).

  4. 4.

    , & Lost after translation: insights from pulmonary surfactant for understanding the role of alveolar epithelial dysfunction and cellular quality control in fibrotic lung disease. Am. J. Physiol. Lung Cell. Mol. Physiol. 309, L507–L525 (2015).

  5. 5.

    , , , & Pulmonary fibrosis in Hermansky–Pudlak syndrome. Ann. Am. Thorac. Soc. 13, 1839–1846 (2016).

  6. 6.

    & Lung development: orchestrating the generation and regeneration of a complex organ. Development 141, 502–513 (2014).

  7. 7.

    & Preparing for the first breath: genetic and cellular mechanisms in lung development. Dev. Cell 18, 8–23 (2010).

  8. 8.

    et al. In vitro generation of human pluripotent stem cell derived lung organoids. eLife 4, (2015).

  9. 9.

    et al. A bioengineered niche promotes in vivo engraftment and maturation of pluripotent stem cell derived human lung organoids. eLife 5, (2016).

  10. 10.

    et al. Efficient generation of lung and airway epithelial cells from human pluripotent stem cells. Nat. Biotechnol. 32, 84–91 (2014).

  11. 11.

    et al. Generation of anterior foregut endoderm from human embryonic and induced pluripotent stem cells. Nat. Biotechnol. 29, 267–272 (2011).

  12. 12.

    et al. The in vitro generation of lung and airway progenitor cells from human pluripotent stem cells. Nat. Protoc. 10, 413–425 (2015).

  13. 13.

    et al. Mesenchymal cells. Defining a mesenchymal progenitor niche at single-cell resolution. Science 346, 1258810 (2014).

  14. 14.

    et al. Hox5 genes regulate the Wnt2/2b-Bmp4-signaling axis during lung development. Cell Rep. 12, 903–912 (2015).

  15. 15.

    et al. Hedgehog signaling in neonatal and adult lung. Am. J. Respir. Cell Mol. Biol. 48, 703–710 (2013).

  16. 16.

    et al. Involvement of Sonic hedgehog (Shh) in mouse embryonic lung growth and morphogenesis. Development 124, 53–63 (1997).

  17. 17.

    , & Sonic hedgehog regulates branching morphogenesis in the mammalian lung. Curr. Biol. 8, 1083–1086 (1998).

  18. 18.

    et al. Multiple dose-dependent roles for Sox2 in the patterning and differentiation of anterior foregut endoderm. Development 134, 2521–2531 (2007).

  19. 19.

    , , , & Two nested developmental waves demarcate a compartment boundary in the mouse lung. Nat. Commun. 5, 3923 (2014).

  20. 20.

    et al. Reconstructing lineage hierarchies of the distal lung epithelium using single-cell RNA-seq. Nature 509, 371–375 (2014).

  21. 21.

    et al. Differential expression of the glycosylated forms of MUC1 during lung development. Eur. J. Histochem. 51, 95–102 (2007).

  22. 22.

    , , , & Recent advances and contraversies on the role of pulmonary neuroepithelial bodies as airway sensors. Semin. Cell Dev. Biol. 24, 40–50 (2013).

  23. 23.

    et al. ABCA3 as a lipid transporter in pulmonary surfactant biogenesis. J. Biol. Chem. 282, 9628–9634 (2007).

  24. 24.

    et al. Life-threatening influenza and impaired interferon amplification in human IRF7 deficiency. Science 348, 448–453 (2015).

  25. 25.

    , & Diseases of pulmonary surfactant homeostasis. Annu. Rev. Pathol. 10, 371–393 (2015).

  26. 26.

    et al. Plasticity of Hopx+ type I alveolar cells to regenerate type II cells in the lung. Nat. Commun. 6, 6727 (2015).

  27. 27.

    & Differential gene expression in the distal tip endoderm of the embryonic mouse lung. Expr. Patterns 2, 229–233 (2002).

  28. 28.

    et al. Sox9 plays multiple roles in the lung epithelium during branching morphogenesis. Proc. Natl Acad. Sci. USA 110, E4456–E4464 (2013).

  29. 29.

    , , , & Normal lung development and function after Sox9 inactivation in the respiratory epithelium. Genesis 41, 23–32 (2005).

  30. 30.

    et al. KeyGenes, a tool to probe tissue differentiation using a human fetal transcriptional atlas. Stem Cell Rep. 4, 1112–1124 (2015).

  31. 31.

    , & Viral bronchiolitis. Lancet 389, 211–224 (2016).

  32. 32.

    et al. Challenges and opportunities in developing respiratory syncytial virus therapeutics. J. Infect. Dis. 211 (suppl. 1), S1–S20 (2015).

  33. 33.

    et al. RSV-encoded NS2 promotes epithelial cell shedding and distal airway obstruction. J. Clin. Invest. 124, 2219–2233 (2014).

  34. 34.

    , , , & Disorders of lysosome-related organelle biogenesis: clinical and molecular genetics. Annu. Rev. Genomics Hum. Genet. 9, 359–386 (2008).

  35. 35.

    et al. Idiopathic pulmonary fibrosis: evolving concepts. Mayo Clin. Proc. 89, 1130–1142 (2014).

  36. 36.

    et al. The alveolar epithelium determines susceptibility to lung fibrosis in Hermansky–Pudlak syndrome. Am. J. Respir. Crit. Care Med. 186, 1014–1024 (2012).

  37. 37.

    Telomerase and idiopathic pulmonary fibrosis. Mutat. Res. 730, 52–58 (2012).

  38. 38.

    , & Spatial mapping and quantification of developmental branching morphogenesis. Development 140, 471–478 (2013).

  39. 39.

    et al. Coordination of heart and lung co-development by a multipotent cardiopulmonary progenitor. Nature 500, 589–592 (2013).

  40. 40.

    , & Generation of three-dimensional lung bud organoid and its derived branching colonies. Protoc. Exch. (2017).

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This work was supported by grant NIH HL120046-01 (H.-W.S.), 1U01HL134760-01 (H.-W.S.) RO1 AI031971 (A.M.), and RO1 AI114736 (A.M.), as well as a sponsored research and agreement from Northern Biologics Inc. (H.-W.S.), and funding from the Thomas R Kully IPF Research Fund (H.-W.S.). RUES2-HPS1 cells were generated by the Columbia Stem Cell Core Facility. We thank NYULMC OCS Microscopy core C. Petzold and K. Dancel for their assistance with transmission electron microscopy. We thank M. Peeples (Ohio State University) for providing the original recombinant RSV. Flow cytometry was performed in the CCTI Flow Cytometry Core, supported in part by the Office of the Director, National Institutes of Health under awards S10RR027050 and S10OD020056.

Author information


  1. Columbia Center for Human Development, Columbia University Medical Center, New York, New York 10032, USA

    • Ya-Wen Chen
    • , Sarah Xuelian Huang
    • , Ana Luisa Rodrigues Toste de Carvalho
    •  & Hans-Willem Snoeck
  2. Columbia Center for Translational Immunology, Columbia University Medical Center, New York, New York 10032, USA

    • Ya-Wen Chen
    • , Sarah Xuelian Huang
    • , Ana Luisa Rodrigues Toste de Carvalho
    • , Siu-Hong Ho
    •  & Hans-Willem Snoeck
  3. Department of Medicine, Columbia University Medical Center, New York, New York 10032, USA

    • Ya-Wen Chen
    • , Sarah Xuelian Huang
    • , Ana Luisa Rodrigues Toste de Carvalho
    • , Siu-Hong Ho
    • , Mohammad Naimul Islam
    • , Jahar Bhattacharya
    •  & Hans-Willem Snoeck
  4. Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, 4710-057 Braga, Portugal

    • Ana Luisa Rodrigues Toste de Carvalho
  5. ICVS/3B’s, PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal

    • Ana Luisa Rodrigues Toste de Carvalho
  6. Division of Immunology and Manton Center for Orphan Disease Research, Children’s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Stefano Volpi
    •  & Luigi D. Notarangelo
  7. U.O. Pediatria 2, Istituto Giannina Gaslini, Genoa 16148, Italy

    • Stefano Volpi
  8. St. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, New York 10065, USA

    • Michael Ciancanelli
    •  & Jean-Laurent Casanova
  9. Department of Physiology & Cellular Biophysics, Columbia University Medical Center, New York, New York 10032, USA

    • Jahar Bhattacharya
    •  & Anne Moscona
  10. OCS Microscopy Core, New York University Langone Medical Center, New York, New York 10016, USA

    • Alice F. Liang
  11. Department of Pediatrics, Columbia University Medical Center, New York, New York 10032, USA

    • Laura M. Palermo
    • , Matteo Porotto
    •  & Anne Moscona
  12. Center for Host-Pathogen Interaction, Columbia University Medical Center, New York, New York 10032, USA

    • Laura M. Palermo
    • , Matteo Porotto
    •  & Anne Moscona
  13. Department of Microbiology and Immunology, Columbia University Medical Center, New York, New York 10032, USA

    • Anne Moscona
    •  & Hans-Willem Snoeck


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Y.-W.C. designed and performed most experiments, contributed to the concept, and co-wrote the manuscript with H.-W.S. S.X.H. and A.L.R.T.d.C. assisted Y.-W.C. A.F.L. performed transmission electron microscopy. S.-H.H. provided assistance with flow cytometry. J.-L.C. and M.C. provided patient material. L.D.N. and S.V. generated the C12 IRF7-deficient iPS line. M.N.I. and J.B. provided SFTPB-BODIPY. A.M., M.P. and L.M.P. generated and provided virology reagents, and provided design and instruction for experiments involving RSV. H.-W.S. provided concept and guidance, and co-wrote with Y.-W.C.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Hans-Willem Snoeck.

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  1. 1.

    Beating cilia in the LBO-derived growth.

    Bright field movie showing beating cilia in LBO-derived growth (RUES2) 5 months after engraftment. Section was made using a vibratome. Scale bar 75 μm.

  2. 2.

    Morphology of d170 LBOs.

    Bright field movie showing the connected dilated saccules and structures resembling pulmonary acini in C12 LBOs. Scale bar 100 μm.

  3. 3.

    Uptake of SPB-BODIPY in d170 LBOs.

    Time lapse movie of several distal buds of different d170 LBOs (RUES2) taken at 2 min intervals. The movie shows uptake of SPB-BODIPY over time in the cells and secretion into the lumen. Green: SPB-BODIPY. Scale bar 50 μm.

  4. 4.

    Modeling respiratory syncytial virus infection in d170 LBOs.

    Confocal microscopy movie of a distal bud of a d170 LBO (RUES2) showing infected cells in the lumen (RSV antigen, green; DAPI, blue). Scale bar 50 μm.

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