Breathing on chip: Dynamic flow and stretch accelerate mucociliary maturation of airway epithelium in vitro .

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 the effects of dynamic medium and airflow as well as stretch on human primary airway epithelial cell differentiation and maturation, including mucociliary clearance, using an organ-on-chip device. Perfused epithelial cell cultures displayed accelerated maturation and polarization of mucociliary clearance, and changes in specific cell-types when compared to traditional (static) culture methods. Additional application of airflow and stretch to the airway chip resulted in an increase in polarization of mucociliary clearance towards the applied flow, 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 maturation and that their fine-tuned application could generate models of specific epithelial pathologies, including mucociliary (dys)function.
Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Anne M. van der Does reports financial support was provided by 10.13039/501100000780European Union. Janna C. Nawroth reports financial support and equipment, drugs, or supplies were provided by Emulate Inc. Doris Roth reports equipment, drugs, or supplies was provided by Emulate Inc. Anne M. van der Does reports equipment, drugs, or supplies was provided by Emulate Inc. Tengku Ibrahim Maulana reports equipment, drugs, or supplies was provided by Emulate Inc. Janna C. Nawroth reports financial support was provided by 10.13039/501100000780European Union. Janna C. Nawroth has patent #US20220106547A1 pending to Emulate Inc. J.N. is a former employee of Emulate, Inc. A.D. was supported by a Global 10.13039/501100000654Marie Curie fellowship (No. 748569) that included a 1-yr visit (2018–2019) at Emulate Inc. to work on their Lung-Chips. D.R. and T.M. performed their internship at Emulate Inc in 2018 and 2019, respectively. Materials from Emulate Inc. Related to this work were therefore provided by Emulate Inc. The remaining authors have no conflicts of interest relevant to this publication.
(© 2023 The Author(s).)