Reproducible uniform equibiaxial stretch of precision-cut lung slices.

Simulating ventilator-induced lung injury (VILI) in the laboratory requires stretching of lung alveolar tissue. Whereas precision-cut lung slices (PCLSs) are widely used for studying paracrine signaling pathways in the lungs, their use in stretch studies is very limited because of the technical challenge of fixing them to a stretchable substrate, stretching them uniformly, or holding them in a stretch device without causing rupture. We describe a novel method for attaching PCLSs to silicone membranes by stitching them together in a star-shaped pattern. Using a device that was previously designed in our laboratory for stretching primary alveolar epithelial cell monolayers, we demonstrate that in the central region of the PCLSs stretch is uniform, equibiaxial, and, after a short preconditioning period, also reproducible. The stitched and stretched PCLSs showed equal or better viability outcomes after 60 min of cyclic stretch at different magnitudes of physiological stretch compared with primary pulmonary alveolar epithelial cell monolayers. Preparing and stitching the PCLSs before stretch is relatively easy to perform, yields repeatable outcomes, and can be used with tissue from any species. Together with the ensuring uniform and equibiaxial stretch, the proposed methods provide an optimal model for VILI studies with PCLSs.