An optimized protocol for generating fibrotic murine precision-cut lung slices.

Area of Science:

• Pulmonary research
• Tissue engineering
• Fibrosis modeling

Background:

• Precision-cut lung slices (PCLSs) are valuable tools in respiratory research, offering advantages over traditional in vivo animal and in vitro cell models.
• Human PCLS availability and ethical considerations can limit their use.
• Need for standardized and reproducible models in pulmonary fibrosis research.

Purpose of the Study:

• To present a detailed protocol for generating fibrotic mouse PCLSs (mPCLSs).
• To establish mPCLSs as a viable alternative to human PCLSs, addressing tissue availability and ethical concerns.
• To enhance experimental homogeneity and reproducibility in pulmonary fibrosis studies.

Main Methods:

• Induction of pulmonary fibrosis in mice using bleomycin (BLM).
• Generation of precision-cut lung slices from fibrotic mouse lungs.
• Visualization and downstream analysis techniques for mPCLSs.

Main Results:

• Successful generation of fibrotic mouse PCLSs (mPCLSs).
• Demonstrated benefits of mPCLSs comparable to human PCLSs.
• Enhanced experimental homogeneity and reproducibility achieved.

Conclusions:

• The developed protocol provides a robust method for creating fibrotic mouse PCLSs.
• mPCLSs serve as an advantageous model for studying pulmonary fibrosis, overcoming limitations of human tissue models.
• This approach supports reproducible and ethically sound respiratory research.