Small airway-on-a-chip enables analysis of human lung inflammation and drug responses in vitro

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Here we describe the development of a human lung 'small airway-on-a-chip' containing a differentiated, mucociliary bronchiolar epithelium and an underlying microvascular endothelium that experiences fluid flow, which allows for analysis of organ-level lung pathophysiology in vitro. Exposure of the epithelium to interleukin-13 (IL-13) reconstituted the goblet cell hyperplasia, cytokine hypersecretion and decreased ciliary function of asthmatics. Small airway chips lined with epithelial cells from individuals with chronic obstructive pulmonary disease recapitulated features of the disease such as selective cytokine hypersecretion, increased neutrophil recruitment and clinical exacerbation by exposure to viral and bacterial infections. With this robust in vitro method for modeling human lung inflammatory disorders, it is possible to detect synergistic effects of lung endothelium and epithelium on cytokine secretion, identify new biomarkers of disease exacerbation and measure responses to anti-inflammatory compounds that inhibit cytokine-induced recruitment of circulating neutrophils under flow.

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Funding was provided by Pfizer, Merck, Wyss Institute for Biologically Inspired Engineering at Harvard University and the Defense Advanced Research Projects Agency (DARPA) under Cooperative Agreement Number W911NF-12-2-0036. We thank K. Karalis for helpful discussions, and B. Hassell and M. Mazur for technical assistance.

Author information

Author notes

    • Carolina Lucchesi
    • , Antonio Varone
    •  & Geraldine A Hamilton

    Present address: Emulate Inc., Cambridge, Massachusetts, USA.

    • Kambez H Benam
    •  & Remi Villenave

    These authors contributed equally to this work.


  1. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA.

    • Kambez H Benam
    • , Remi Villenave
    • , Carolina Lucchesi
    • , Antonio Varone
    • , Thomas C Ferrante
    • , James C Weaver
    • , Anthony Bahinski
    • , Geraldine A Hamilton
    •  & Donald E Ingber
  2. Pfizer, Cambridge, Massachusetts, USA.

    • Cedric Hubeau
  3. Merck Research Laboratories, Boston, Massachusetts, USA.

    • Hyun-Hee Lee
    • , Stephen E Alves
    •  & Michael Salmon
  4. Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA.

    • James C Weaver
    •  & Donald E Ingber
  5. Vascular Biology Program, Boston Children's Hospital, Boston, Massachusetts, USA.

    • Donald E Ingber
  6. Harvard Medical School, Harvard University, Boston, Massachusetts, USA.

    • Donald E Ingber


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K.H.B., R.V., C.H., H.-H.L., S.E.A., M.S., G.A.H. and D.E.I. designed the research; K.H.B. and R.V. developed the basic small airway chip model; R.V. and A.V. conducted the asthma work; K.H.B. and C.L. conducted the COPD studies; K.H.B. optimized and performed leukocyte-recruitment studies; J.C.W. performed scanning electron microscopy imaging; T.C.F. helped with confocal microscopy imaging; K.H.B. and R.V. prepared the manuscript; G.A.H. and A.B. commented on the manuscript; and D.E.I. critically revised the manuscript.

Competing interests

D.E.I. and G.A.H. are founders and hold equity in Emulate, Inc., and D.E.I. chairs its scientific advisory board.

Corresponding author

Correspondence to Donald E Ingber.

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Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–3, Supplementary Tables 1 and 2, and Supplementary Discussion


  1. 1.

    3D visualization of the lung small airway epithelium and endothelium reconstituted on-chip.

    A movie showing various confocal fluorescence microscopic 3D views of a fully differentiated, pseudostratified, human small airway epithelium cultured at an air-liquid interface, formed from primary hAECs (F-actin, green) and cocultured with primary human pulmonary microvascular endothelial cells (F-actin, red) in the top and lower channels of the small airway chip device, respectively (DAPI-stained nuclei, blue).

  2. 2.

    Z-stack reconstructed 3D visualization of the mucociliary small airway epithelium on-chip.

    A video showing multiple confocal immunofluorescence 3D views of a polarized, mucociliary, human bronchiolar epithelium grown in the small airway chip showing cilia stained with anti−β-tubulin (cyan) and goblet cells labeled with anti-MUC5AC (magenta).

  3. 3.

    Active ciliary beating of the differentiated human airway epithelium on-chip.

    Time-lapse video of phase-contrast views of the human small airway epithelium with apical cilia beating actively on-chip. The video is slowed down to enable analysis of cilia beating frequencies in the region of interest (square at top left).

  4. 4.

    Visualization of human airway epithelial mucociliary transport on-chip.

    Real-time fluorescence microscopic imaging of mucociliary transport by the differentiated human airway epithelium cultured on-chip when exposed to fluorescent 1-μm-diameter microbeads (white; scale bar, 50 μm).

  5. 5.

    Recruitment and adhesion of circulating human leukocytes in the human small airway chip.

    Real-time fluorescence microscopic imaging showing how freshly isolated, CellTracker red−labeled human neutrophils adhere to the endothelium and roll over its surface (a single adherent neutrophil is shown in this high-magnification view) when these leukocytes are flowed under physiological conditions (shear stress, 1 dyn cm−2) through the microvascular (endothelium-lined) channel of a human small airway chip that was stimulated by the addition of poly(I:C) to the epithelium in its upper channel.