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Protocol for isolation and flow cytometric analysis of interstitial macrophages in murine lung metastasis models

Year: 2025
Type: article
Abstract: Interstitial macrophages increase significantly during lung metastasis and may contribute to tumor dissemination. However, isolating them is challenging due to their localization within lung tissue and phenotypic overlap with other immune cells. Here, we present a protocol for isolating and characterizing murine interstitial macrophages. We include steps for establishing lung metastasis models, lung tissue dissociation, fluorescent antibody staining, and flow cytometry analysis. It is applicable to both metastasized and naive lung tissues. For complete details on the use and execution of this protocol, please refer to Jeong et al. 1 • Steps for generating mouse lung metastasis models using cancer cell lines • Procedures for isolating interstitial macrophages from metastasized lungs • Guidance for gating lung interstitial macrophages for flow cytometry Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Interstitial macrophages increase significantly during lung metastasis and may contribute to tumor dissemination. However, isolating them is challenging due to their localization within lung tissue and phenotypic overlap with other immune cells. Here, we present a protocol for isolating and characterizing murine interstitial macrophages. We include steps for establishing lung metastasis models, lung tissue dissociation, fluorescent antibody staining, and flow cytometry analysis. It is applicable to both metastasized and naive lung tissues.
DOI: 10.1016/j.xpro.2025.104060
Language: English
Subfield: Immunology
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W4414042064
FWCI: 0.303
Cites: 6
Cited by: 1
Related to: 10