Phenotypic Characterization and Isolation of Myeloid-Derived Suppressor Cells
Or Reuven1, Ivan Mikula1, Hadas Ashkenazi-Preiser1
1The Concern Foundation Laboratories at The Lautenberg Center for Immunology and Cancer Research, Israel-Canada Medical Research Institute, Faculty of Medicine, The Hebrew University, Jerusalem, Israel.
Current Protocols
|October 10, 2022
Summary
This study details methods for isolating and characterizing myeloid-derived suppressor cells (MDSCs) and their subpopulations from mouse tissues and human blood. These protocols enable the study of MDSCs in various diseases, including cancer and autoimmune conditions.
Area of Science:
- Immunology
- Cell Biology
- Cancer Research
Background:
- Myeloid-derived suppressor cells (MDSCs) are immature myeloid cells that suppress immune responses.
- MDSCs comprise polymorphonuclear (PMN) and monocytic (M) subpopulations.
- These cells are implicated in pathologies like cancer, inflammatory bowel disease, and autoimmune disorders.
Purpose of the Study:
- To provide standardized methods for isolating and characterizing MDSCs.
- To enable the study of MDSC distribution, maturation, and phenotype.
- To facilitate research on MDSCs in both murine models and human patients.
Main Methods:
- Isolation of total MDSCs and subpopulations (PMN-MDSCs, M-MDSCs) from murine tissues.
- Phenotyping of MDSCs using flow cytometry, immunofluorescence, and Giemsa staining.
- Protocols for handling human blood samples and characterizing blood-derived MDSCs.
Main Results:
- Established protocols for generating single-cell suspensions from various tissues, including tumors.
- Detailed methods for cell separation using magnetic beads and for flow cytometry and immunofluorescence analyses.
- Provided a gating strategy for PMN-MDSCs and M-MDSCs in mice.
Conclusions:
- The described methods offer a comprehensive approach to MDSC isolation and characterization.
- These protocols are applicable to both preclinical research and clinical sample analysis.
- Standardized methods are crucial for advancing the understanding and therapeutic targeting of MDSCs.
