ArticleCurrent protocols2025
Analyzing Pathophysiology and Immune Cells and Their Cytokines and Mediators in Precision-Cut Slices of the Murine Lung.
Article in Current protocols, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- Immunoregulatory role of quercetin in lung cancer.Cell death discovery · 2026Article
- Preparation and Infection of Bovine Precision-Cut Lung Slices (b-PCLS) as an Ex Vivo Model for Respiratory Pathogens.Current protocols · 2026Article
- Comprehensive analysis of diverse cytokine patterns in the prognosis and tumor microenvironment of lung adenocarcinoma.Discover oncology · 2026Article
- Analyzing Pathophysiology and Immune Cells and Their Cytokines and Mediators in Precision-Cut Slices of the Murine Lung.Current protocols · 2025Article
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9 authors.
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Abstract
Understanding the dynamic pathophysiology of diseases in the lung, such as asthma and chronic asthma, chronic obstructive pulmonary disease, and lung cancer, is crucial for the treatment, analysis, and outcome of these diseases. Unlike other traditional models, we suggest a protocol that is sustainable and reproducible and offers different analysis methods while maintaining in vivo lung architecture and immune dynamics. This protocol allows one to study the pathophysiological changes, including changes to the immune cells, cytokines, and mediators, in 30 precision-cut lung slices from a single murine lung. To accomplish this, the murine lung is infused with 2.5% low-melting-point agarose and is precision-cut-sliced. Our method also supports cell culture in refined medium and stimulation with clinically relevant stimuli, which helps to clarify the mechanisms of the disease. Evaluation of the samples and their supernatant includes multiplex assays, ELISA, histology, and immunohistochemistry. Additional sections are used to extract RNA for quantitative real-time PCR and RNA sequencing and/or other selected analysis, like flow cytometry. Using this method, we obtained murine lung slices that preserve the pathophysiology of the disease and allow a comprehensive analysis unlike other already-existing protocols. By retaining the dynamic immune mechanisms, we are able to see the histological damage caused by each disease. The results of this protocol can be used to improve our understanding and therapy options. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Obtaining precision-cut lung slices (PCLSs) from the murine lung Support Protocol 1: LL/2 cell treatment with G-418 solution Support Protocol 2: Murine model of lung adenocarcinoma and in vivo imaging Support Protocol 3: H&E staining of PCLS sections Support Protocol 4: Measuring cytokines by ELISA Support Protocol 5: Measuring cytotoxic activity in PCLS conditioned medium Support Protocol 6: Analysis of RNA from PCLSs by real-time PCR Support Protocol 7: Flow cytometry analysis of cells isolated from PCLSs.
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