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Lipid-Activated Nuclear Receptors pp 59–73Cite as

Isolation and Purification of Tissue Resident Macrophages for the Analysis of Nuclear Receptor Activity

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1951))

Abstract

Tissue resident macrophages (TRMs) are multifunctional immune cells present in all tissues, contributing to the correct development, homeostasis, and protection against pathogens and injury. TRMs are morphologically and functionally heterogeneous, as a result of both the diversity of tissue environments in which they reside and their complex origin. Furthermore, some specific TRM populations are controlled by nuclear receptors. A widely used method for studying the role of nuclear receptors in immune cells is flow cytometry. Although flow cytometry is extensively used in tissues such as the peripheral blood, lymph nodes, peritoneal cavity, and bone marrow, there is a need for protocols for the study TRMs in solid tissues.

In this chapter, we describe a comprehensive protocol for obtaining single-cell suspensions of resident macrophages from the pleural cavity, heart, lung, spleen, and kidney, and we present detailed gating strategies for the study of nuclear receptor activity in different TRM subsets within these tissues.

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Acknowledgment

We thank Simon Bartlett for English editing. This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2015-64287R, SAF2017-90604-REDT), Fundació Marató TV3 (121931), and the Community of Madrid (B2017/BMD-3684) to M.R. L.A-H. is funded by a fellowship from Obra Social “La Caixa”. The CNIC is supported by the MEIC and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence awardee (MEIC award SEV-2015-0505).

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Author notes

  1. Laura Alonso-Herranz and Jesús Porcuna contributed equally to this work.

Authors and Affiliations

  1. Myocardial Pathophysiology Area, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Madrid, Spain

    Laura Alonso-Herranz, Jesús Porcuna & Mercedes Ricote

Authors
  1. Laura Alonso-Herranz
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  2. Jesús Porcuna
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  3. Mercedes Ricote
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Corresponding author

Correspondence to Mercedes Ricote .

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Editors and Affiliations

  1. Division of Medicine, Centre for Cardiometabolic Medicine, University College London, London, UK

    Matthew C. Gage

  2. Division of Medicine, Centre for Cardiometabolic Medicine, University College London, London, UK

    Inés Pineda-Torra

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Alonso-Herranz, L., Porcuna, J., Ricote, M. (2019). Isolation and Purification of Tissue Resident Macrophages for the Analysis of Nuclear Receptor Activity. In: Gage, M., Pineda-Torra, I. (eds) Lipid-Activated Nuclear Receptors. Methods in Molecular Biology, vol 1951. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9130-3_5

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  • DOI: https://doi.org/10.1007/978-1-4939-9130-3_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9129-7

  • Online ISBN: 978-1-4939-9130-3

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