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Picturing Polarized Myeloid Phagocytes and Regulatory Cells by Mass Cytometry

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Book cover Mass Cytometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1989))

Abstract

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.

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Author information

Authors and Affiliations

  1. Univ Rennes, CHU Rennes, Inserm, MICMAC [(MIcroenvironment, Cell differentiation, iMmunology And Cancer)]—UMR_S 1236, EFS, Rennes, France

    Mikael Roussel

  2. Department of Pathology, Microbiology and Immunology and Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN, USA

    Todd Bartkowiak & Jonathan M. Irish

  3. Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    Todd Bartkowiak & Jonathan M. Irish

Authors
  1. Mikael Roussel
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  2. Todd Bartkowiak
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  3. Jonathan M. Irish
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Corresponding authors

Correspondence to Mikael Roussel or Jonathan M. Irish .

Editor information

Editors and Affiliations

  1. Discipline of Pathology, The University of Sydney, Camperdown, NSW, Australia

    Helen M. McGuire

  2. Sydney Cytometry Facility, The University of Sydney and Centenary Institute, Camperdown, NSW, Australia

    Thomas M. Ashhurst

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Roussel, M., Bartkowiak, T., Irish, J.M. (2019). Picturing Polarized Myeloid Phagocytes and Regulatory Cells by Mass Cytometry. In: McGuire, H., Ashhurst, T. (eds) Mass Cytometry. Methods in Molecular Biology, vol 1989. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9454-0_14

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  • DOI: https://doi.org/10.1007/978-1-4939-9454-0_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9453-3

  • Online ISBN: 978-1-4939-9454-0

  • eBook Packages: Springer Protocols

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