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Enrichment and Characterization of the Tumor Immune and Non-immune Microenvironments in Established Subcutaneous Murine Tumors
Published on: June 7, 2018
Murine brain tumor microenvironment immunophenotyping using mass cytometry
Brandon L McClellan1,2, Mahmoud S Alghamri1,2, Rohit Thalla1,2
1Department of Neurosurgery, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Insights
This study details a mass cytometry protocol for analyzing immune cells in mouse glioma. The optimized method aids in understanding the tumor microenvironment in brain cancer research.
Area of Science:
- Immunology
- Oncology
- Neuroscience
Background:
- The glioma microenvironment harbors diverse immune cells crucial for tumor progression.
- Understanding immune cell phenotypes is vital for developing effective brain cancer therapies.
Purpose of the Study:
- To present an optimized mass cytometry protocol for dissecting immune cell phenotypes in mouse glioma models.
- To provide a comprehensive guide for antibody conjugation, tissue processing, and data analysis.
Main Methods:
- Utilized a Sleeping Beauty transposon-mediated mouse glioma model.
- Developed and optimized antibody conjugation and titration strategies.
- Detailed procedures for brain tumor tissue collection, processing, staining, data acquisition, and analysis.
Main Results:
- Established a robust mass cytometry protocol applicable to various brain tumor mouse models.
- Enabled detailed phenotypic analysis of immune cells within the glioma microenvironment.
Conclusions:
- The presented protocol offers a standardized approach for immune cell profiling in brain tumors.
- Facilitates in-depth investigation of the immune landscape in glioma research.
Abstract:
Here, we present a mass cytometry protocol optimized to examine the phenotype of immune cells within the mouse glioma microenvironment, using a Sleeping Beauty transposon-mediated mouse glioma model. We describe antibody conjugation and titrations for analysis of immune cells. We then detail mouse brain tumor tissue collection and processing, staining, followed by data acquisition, analysis, and gating strategy. This protocol can be applied to any brain tumor-harboring mouse model. For complete details on the use and execution of this protocol, please refer to Alghamri et al. (2021).
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