Mass Cytometry pp 217-226 | Cite as

Picturing Polarized Myeloid Phagocytes and Regulatory Cells by Mass Cytometry

  • Mikael RousselEmail author
  • Todd Bartkowiak
  • Jonathan M. IrishEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1989)


The immune monocyte/phagocyte system (MPS) includes numerous cell subsets of the myeloid lineage including monocyte, macrophage, and dendritic cell (DC) populations that are heterogeneous both phenotypically and functionally. Previously, we characterized these diverse MPS phenotypes with multi-parametric mass cytometry (CyTOF). In order to expansively characterize monocytes, macrophages, and dendritic cells, a CyTOF panel was designed to measure 35 identity-, activation-, and polarization-markers. Here we provide a protocol to define a reference map for the myeloid compartment, including sample preparation, to produce reference cell subsets from the monocyte/phagocyte system. In particular, we focused on monocyte-derived macrophages that were further polarized in vitro with cytokine stimulation (i.e., M-CSF, GM-CSF, IL-4, IL-10, IFNγ, and LPS), as well as monocyte-derived DCs, and myeloid-derived suppressor cells (MDSCs), generated in vitro from human bone marrow and/or peripheral blood.

Key words

Mass cytometry Macrophage polarization Dendritic cells Myeloid-derived suppressor cells Myeloid regulatory cells 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Univ Rennes, CHU Rennes, Inserm, MICMAC [(MIcroenvironment, Cell differentiation, iMmunology And Cancer)]—UMR_S 1236, EFSRennesFrance
  2. 2.Department of Pathology, Microbiology and Immunology and Vanderbilt-Ingram Cancer CenterVanderbilt University Medical CenterNashvilleUSA
  3. 3.Department of Cell and Developmental BiologyVanderbilt University School of MedicineNashvilleUSA

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