An Ex Vivo Acid Injury and Repair (AIR) Model Using Precision-Cut Lung Slices to Understand Lung Injury and Repair

Area of Science:

• Pulmonary Medicine
• Cell Biology
• Toxicology

Background:

• Current lung cell culture models do not fully replicate the lung's complex 3D environment.
• Precision-cut lung slices (PCLS) offer a more physiologically relevant ex vivo model.
• A need exists for robust models to study acute lung injury and repair.

Purpose of the Study:

• To present detailed protocols for generating PCLS and the novel acid injury and repair (AIR) model.
• To establish a reproducible ex vivo method for studying lung injury and repair.
• To enable semi-high-throughput screening of potential therapeutics for lung injury.

Main Methods:

• Generation of precision-cut lung slices (PCLS) from mouse lungs.
• Induction of spatially restricted tissue injury using HCl and Pluronic gel (AIR model).
• Assessment of injury and repair via immunofluorescence for markers like Ki67 and prosurfactant protein C.

Main Results:

• The AIR model allows ex vivo study of lung injury and repair without prior in vivo injury.
• Immunofluorescence staining with established markers effectively quantifies injury and repair.
• The AIR model demonstrates high reproducibility in assessing lung tissue responses.

Conclusions:

• The AIR model using PCLS is a versatile tool for investigating lung cell biology in acute lung injury.
• This ex vivo approach bridges the gap between in vitro and in vivo lung models.
• The AIR model facilitates the screening of potential therapeutic compounds for lung injury.