A lung-on-a-chip array with an integrated bio-inspired respiration mechanism

Abstract

We report a lung-on-a-chip array that mimics the pulmonary parenchymal environment, including the thin alveolar barrier and the three-dimensional cyclic strain induced by breathing movements. The micro-diaphragm used to stretch the alveolar barrier is inspired by the in vivo diaphragm, the main muscle responsible for inspiration. The design of this device aims not only at best reproducing the in vivo conditions found in the lung parenchyma but also at making the device robust and its handling easy. An innovative concept, based on the reversible bonding of the device, is presented that enables accurate control of the concentration of cells cultured on the membrane by easily accessing both sides of the membranes. The functionality of the alveolar barrier could be restored by co-culturing epithelial and endothelial cells that form tight monolayers on each side of a thin, porous and stretchable membrane. We showed that cyclic stretch significantly affects the permeability properties of epithelial cell layers. Furthermore, we also demonstrated that the strain influences the metabolic activity and the cytokine secretion of primary human pulmonary alveolar epithelial cells obtained from patients. These results demonstrate the potential of this device and confirm the importance of the mechanical strain induced by breathing in pulmonary research.

Human lung epithelial cell cultures for analysis of inhaled toxicants: Lessons learned and future directions

“…The relevance of these findings on the importance of cycling breathing was confirmed in an ex vivo mouse ventilation perfusion model. These findings on the importance of mechanical stretch for cellular behaviour in an alveolus-ona-chip model was confirmed and extended to include metabolic activity and cytokine production by other groups (Stucki et al, 2015). Furthermore, the group of Ingber at the Wyss Institute reported on application of their model for studies on IL-2-induced pulmonary oedema, P.S.…”

Towards personalized medicine: chemosensitivity assays of patient lung cancer cell spheroids in a perfused microfluidic platform

“…The appropriate amount of cells was then seeded either on chip for perfused experiments or in a 96 U-bottom well plate (BD Falcon) for static experiments in RPMI1640 medium supplemented 20 ng/ml of human epi- View Article Online dermal growth factor, hEGF (Gibco, Invitrogen), 20 ng/ml of basic fibroblast growth factor, bFGF (Gibco, Invitrogen), 4 µg/ml of human insulin (BioReagent, Sigma), 2% serum-free supplement B27 (Gibco, Invitrogen) and 1% P/S. Primary lung epithelial tumor cells (PLETCs) as well as primary pericytes (PCs) were isolated from lung tumor specimens as previously described 26 . In brief, PLETCs were prospectively isolated using fluorescent activated cell sorting (FACS) with an immunophenotypic profile of LineageEpCAM+CD73+CD90-, and seed for expansion in a 6 well dish coated with 0.2% gelatin and human collagen IV (Sigma) in CnT-PA growth medium (CellnTec, Switzerland) supplemented with 10 ng/ml insulin like growth factor, IGF-2 (Peprotech) and 10 ng/ml Heregulin β , HerB (Peprotech).…”

Non-animal models of epithelial barriers (skin, intestine and lung) in research, industrial applications and regulatory toxicology

“…By using electrical impedance, the electrical barrier properties of epithelial cells can be monitored continuously without submerging the cultures (Sun et al, 2010). A more advanced model of the pulmonary epithelium is the so called "lung-on-a-chip" model, a device with alveolar epithelial cells on the apical and endothelial cells on the basolateral side of a porous membrane (Huh et al, 2010;Stucki et al, 2015). In this model, the cells are supplied with nutrition via microfluidic flow of medium which also allows introduction of immune cells into the model.…”