Protocol for the establishment of murine orthotopic lung tumors and the assessment of contralateral pulmonal metastasis by flow cytometry
Murine orthotopic tumor models can accurately resemble cancer biology characteristics including metastasis, drug sensitivity, and remodeling of the tumor microenvironment. Here, we present a protocol for establishing murine orthotopic lung tumors and assessing contralateral pulmonary metastasis by flow cytometry. We describe steps for marking tumor cells with label retention dyes, preparing cell mixtures in Matrigel, and performing intrapulmonary injection. We then detail procedures for ex vivo processing of lungs followed by fluorescence-activated cell sorting (FACS) data analysis and quantification of metastatic capacity. For complete details on the use and execution of this protocol, please refer to Kalkavan et al.<sup>1</sup>.
Authors
- Gregor Zaun (ORCID: https://orcid.org/0000-0002-3813-6961)
- Diego Rodriguez (ORCID: https://orcid.org/0000-0002-8055-9511)
- Maximilian Webendörfer (ORCID: https://orcid.org/0009-0006-1498-3134)
- Halime Kalkavan (ORCID: https://orcid.org/0000-0002-0090-4080)
- Douglas R. Green (ORCID: https://orcid.org/0000-0002-7332-1417)
- Si‐Yang Maggie Liu (ORCID: https://orcid.org/0000-0002-3837-0758)
- Abubakar Wani (ORCID: https://orcid.org/0000-0001-7079-3278)
Institutions
- St. Jude Children's Research Hospital (US)
- University Hospital Bonn (DE)
- Essen University Hospital (DE)
- West German Heart and Vascular Center Essen (DE)
- University of Duisburg-Essen (DE)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-04-23
- DOI
- https://doi.org/10.1016/j.xpro.2025.103793
- Citations
- 1
- Primary Topic
- Neonatal Respiratory Health Research
- Type
- article
- Field-Weighted Citation Impact
- 1.52
Funders
- National Foundation for Cancer Research
- American Lebanese Syrian Associated Charities
- Deutsche Forschungsgemeinschaft
- Bundesministerium für Bildung und Forschung
- Deutsche Krebshilfe
- Universität Duisburg-Essen
- Bundesministerium für Bildung, Wissenschaft, Forschung und Technologie
- National Cancer Institute