Phenotypic Characterization and Isolation of Myeloid-Derived Suppressor Cells

Or Reuven, Ivan Mikula, Hadas Ashkenazi-Preiser, Nira Twaik, Kerem Ben-Meir, Yaron Meirow, Leonor Daniel, Guy Kariv, Mahdi Kurd, Michal Baniyash

Research output: Contribution to journalArticlepeer-review


Myeloid-derived suppressor cells (MDSCs) are heterogenous populations of immature myeloid cells that can be divided into two main subpopulations, polymorphonuclear (PMN) MDSCs and monocytic (M) MDSCs. These cells accumulate during chronic inflammation and induce immunosuppression evident in an array of pathologies such as cancer, inflammatory bowel disease, and infectious and autoimmune diseases. Herein, we describe methods to isolate and characterize MDSCs from various murine tissue, as well as to phenotype blood-derived MDSCs from patients. The protocols describe methods for isolation of total MDSCs and their subpopulations, for characterization, and for evaluation of their distribution within tissue, as well as for assessing their maturation stage by flow cytometry, immunofluorescence analyses, and Giemsa staining. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Single-cell suspension generation from different tissue Alternate Protocol 1: Single-cell suspension generation from subcutaneous melanoma tumors Basic Protocol 2: Characterization of MDSC phenotype Basic Protocol 3: Cell separation using magnetic beads: Separating pan-MDSCs or PMN-MDSC and M-MDSC subpopulations Alternate Protocol 2: Staining and preparing MDSCs for sorting Support Protocol: PMN-MDSC and M-MDSC gating strategy in mouse Basic Protocol 4: Immunofluorescence analysis of MDSCs Basic Protocol 5: Handling human blood samples and characterizing human MDSCs Alternate Protocol 3: Flow cytometry staining of thawed human whole blood samples.

Original languageAmerican English
Article numbere561
JournalCurrent Protocols
Issue number10
StatePublished - Oct 2022


  • MDSC
  • chronic inflammation
  • flow cytometry
  • immunofluorescence imaging
  • immunosuppression
  • inflammatory mouse model
  • myeloid-derived suppressor cells
  • patient MDSC

All Science Journal Classification (ASJC) codes

  • General Immunology and Microbiology
  • General Pharmacology, Toxicology and Pharmaceutics
  • Medical Laboratory Technology
  • General Biochemistry,Genetics and Molecular Biology
  • Health Informatics
  • General Neuroscience


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