Breathing on Chip: Dynamic flow and stretch tune cellular composition and accelerate mucociliary maturation of airway epithelium in vitro

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract Human lung function is intricately linked to blood flow and breathing cycles, but it remains unknown how these dynamic cues shape human airway epithelial biology. Here we report a state-of-the-art protocol for studying effects of physiological airflow and stretch on differentiation, cellular composition, and mucociliary clearance of human primary airway epithelial cells cultured on a perfused airway chip. Perfused epithelial tissue cultures developed a large airway-like cellular composition with accelerated maturation and polarization of mucociliary clearance when compared to traditional (static) culture methods. Additional application of airflow and stretch to the airway chip resulted in a cellular composition more comparable to the small(er) airways, reduced baseline secretion of interleukin-8 and other inflammatory proteins, and reduced gene expression of matrix metalloproteinase (MMP) 9, fibronectin, and other extracellular matrix factors. These results indicate that breathing-like mechanical stimuli are important modulators of airway epithelial cell differentiation and homeostasis and that their fine-tuned application could generate models of specific epithelial regions, pathologies, and mucociliary (dys)function.

Authors 12

1. Janna C. Nawroth

   University of Southern California · Emulate (United States)

   Affiliation as printed

   Emulate Inc., Boston, MA, U.S.A

   Hastings Center for Pulmonary Research, Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, University of Southern California, Los Angeles, CA, U.S.A

2. Annemarie M. van Schadewijk Aachen Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Dept. of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

3. Abilash Ravi Aachen Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Dept. of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

4. Tengku Ibrahim Maulana

   Emulate (United States)

   Affiliation as printed

   Emulate Inc., Boston, MA, U.S.A

5. Christiana N. Senger

   University of Southern California

   Affiliation as printed

   Hastings Center for Pulmonary Research, Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, University of Southern California, Los Angeles, CA, U.S.A

6. Sander van Riet Aachen Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Dept. of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

7. Dennis K. Ninaber Aachen Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   Dept. of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

8. Amy M. de Waal
9. Dorothea Kraft
10. Pieter Sicco Hiemstra Aachen Dept of Pulmonology, Leiden University Medical Center

    Leiden University Medical Center

    Affiliation as printed

    Dept. of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

11. Amy L. Ryan

    University of Southern California

    Affiliation as printed

    Department of Stem Cells and Regenerative Medicine, University of Southern California, Los Angeles, CA, U.S.A

    Hastings Center for Pulmonary Research, Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, University of Southern California, Los Angeles, CA, U.S.A

12. Anne M. van der Does corresponding Aachen Dept of Pulmonology, Leiden University Medical Center

    Leiden University Medical Center

    Affiliation as printed

    Dept. of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands