Reproducible uniform equibiaxial stretch of precision-cut lung slices.

Area of Science:

• Biomedical Engineering
• Pulmonary Medicine
• Cell Biology

Background:

• Simulating ventilator-induced lung injury (VILI) requires stretching lung alveolar tissue.
• Precision-cut lung slices (PCLSs) are valuable for lung research but difficult to use in stretch studies due to technical challenges.
• Existing methods struggle with uniform stretching and preventing tissue rupture.

Purpose of the Study:

• To develop a novel, reliable method for stretching PCLSs.
• To enable uniform and reproducible mechanical stretch of PCLSs for VILI research.
• To improve the viability and applicability of PCLSs in mechanical stretch studies.

Main Methods:

• A new technique was developed to attach PCLSs to silicone membranes using a star-shaped stitching pattern.
• A previously designed stretching device for cell monolayers was adapted for the stitched PCLSs.
• PCLSs were subjected to cyclic stretch at various physiological magnitudes to assess uniformity, reproducibility, and viability.

Main Results:

• The novel stitching method allowed for uniform, equibiaxial, and reproducible stretch in the central region of PCLSs.
• Stitched and stretched PCLSs demonstrated equal or superior viability compared to primary alveolar epithelial cell monolayers after 60 minutes of cyclic stretch.
• The method is easy to perform, yields repeatable outcomes, and is applicable to lung tissue from any species.

Conclusions:

• The proposed method provides an optimal and robust model for VILI studies using PCLSs.
• This technique overcomes previous limitations in stretching lung tissue slices.
• It facilitates advanced research into the mechanisms of VILI and potential therapeutic interventions.