Immunohistochemical Identification of Human Skeletal Muscle Macrophages

Bio-protocol
Kate KosmacCharlotte A Peterson

Abstract

Macrophages have well-characterized roles in skeletal muscle repair and regeneration. Relatively little is known regarding the role of resident macrophages in skeletal muscle homeostasis, extracellular matrix remodeling, growth, metabolism and adaptation to various stimuli including exercise and training. Despite speculation into macrophage contributions during these processes, studies characterizing macrophages in non-injured muscle are limited and methods used to identify macrophages vary. A standardized method for the identification of human resident skeletal muscle macrophages will aide in the characterization of these immune cells and allow for the comparison of results across studies. Here, we present an immunohistochemistry (IHC) protocol, validated by flow cytometry, to distinctly identify resident human skeletal muscle macrophage populations. We show that CD11b and CD206 double IHC effectively identifies macrophages in human skeletal muscle. Furthermore, the majority of macrophages in non-injured human skeletal muscle show a 'mixed' M1/M2 phenotype, expressing CD11b, CD14, CD68, CD86 and CD206. A relatively small population of CD11b+/CD206- macrophages are present in resting skeletal muscle. Changes in the relative abu...Continue Reading

References

Sep 25, 2001·Journal of Applied Physiology·N StupkaS M Phillips
Nov 2, 2002·The Journal of Physiology·Louise J BeatonStuart M Phillips
Jun 5, 2003·Medicine and Science in Sports and Exercise·Jennifer M PetersonFrancis X Pizza
Aug 7, 2004·Matrix Biology : Journal of the International Society for Matrix Biology·Regina M CrameriMichael Kjaer
Jul 26, 2006·The Journal of Cell Biology·Katie M Jansen, Grace K Pavlath
Nov 4, 2006·The Journal of Immunology : Official Journal of the American Association of Immunologists·Fernando O MartinezAlberto Mantovani
May 9, 2007·The Journal of Experimental Medicine·Ludovic ArnoldBénédicte Chazaud
Jul 27, 2007·American Journal of Physiology. Cell Physiology·Dana M DiPasqualeTimothy J Koh
Mar 7, 2008·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·D J MahoneyM A Tarnopolsky
Apr 15, 2008·Scandinavian Journal of Immunology·E GottfriedM Kreutz
Oct 22, 2008·Sports Medicine·Carine SmithKathryn H Myburgh
Nov 26, 2008·Nature Reviews. Immunology·David M Mosser, Justin P Edwards
Dec 23, 2008·Exercise and Sport Sciences Reviews·Bénédicte ChazaudFabrice Chretien
Apr 2, 2009·American Journal of Physiology. Endocrinology and Metabolism·Vijayalakshmi VarmaCharlotte A Peterson
Jun 16, 2009·Scandinavian Journal of Medicine & Science in Sports·G PaulsenT Raastad
Dec 17, 2009·Medicine and Science in Sports and Exercise·Gøran PaulsenTruls Raastad
Mar 12, 2010·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·James G Tidball, S Armando Villalta
Dec 15, 2010·CNS & Neurological Disorders Drug Targets·M OlahH W G M Boddeke
Mar 4, 2011·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Lauren G MacNeilMark A Tarnopolsky
Apr 5, 2011·Muscle & Nerve·Mark A TarnopolskyBoleslaw Lach
Nov 15, 2011·Journal of Immunological Methods·C A AmbarusD L P Baeten
Feb 7, 2013·The FEBS Journal·Marielle SaclierBénédicte Chazaud
Mar 20, 2013·Mediators of Inflammation·Yacine KharrazPura Muñoz-Cánoves
Oct 5, 2013·The American Journal of Pathology·Margaret L Novak, Timothy J Koh
Oct 9, 2013·Proceedings of the National Academy of Sciences of the United States of America·Frances Y McWhorterWendy F Liu
Oct 15, 2013·Biochemical and Biophysical Research Communications·Shin-ichi IkedaHirotaka Watada
Feb 15, 2014·The American Journal of Pathology·Hanzhou WangPaula K Shireman
Mar 7, 2014·Development·James G TidballMichelle Wehling-Henricks
Mar 29, 2014·F1000prime Reports·Fernando O Martinez, Siamon Gordon
May 27, 2014·BioMed Research International·Elena RigamontiSilvia Brunelli
Oct 7, 2014·Journal of Visualized Experiments : JoVE·R Andrew ShanelyDavid C Nieman
Oct 17, 2014·Immunological Reviews·Siamon GordonFernando Martinez Estrada
Nov 5, 2014·Frontiers in Immunology·Paola Italiani, Diana Boraschi
Sep 25, 2015·Nature Protocols·Ling LiuThomas A Rando
Feb 13, 2016·Cell Death and Differentiation·C ScioratiP Rovere-Querini

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Citations

Apr 7, 2019·Cell and Tissue Research·Douglas Florindo PinheiroJussara Lagrota-Candido
Jul 26, 2019·Journal of Applied Physiology·Jonathan B MuyskensHans C Dreyer
Aug 10, 2019·Stem Cells International·Junio DortNicolas A Dumont
Sep 4, 2019·Aging Cell·Chang-Yi CuiLuigi Ferrucci
Jun 19, 2019·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Jacob R SorensenRobert D Hyldahl
Feb 14, 2020·Journal of Applied Physiology·Muthita Hirunsai, Ratchakrit Srikuea
Mar 13, 2020·The Journal of Immunology : Official Journal of the American Association of Immunologists·David M StepienBenjamin Levi
May 12, 2020·Journal of the American Heart Association·Kate KosmacCharlotte A Peterson
Aug 29, 2020·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Masayuki MiyagiMasashi Takaso
May 30, 2020·Applied Physiology, Nutrition, and Metabolism = Physiologie Appliquée, Nutrition Et Métabolisme·Paul T ReidyMicah J Drummond
Mar 3, 2020·Frontiers in Immunology·Chanju LeeHyunsu Bae
Dec 6, 2019·Journal of Microscopy and Ultrastructure·Faizah AlotaibiHeba Fikry
Jul 1, 2020·Cells·Anastasiya S PoltavetsTimur Kh Fatkhudinov
Nov 13, 2020·Frontiers in Physiology·Patricia S RogeriAntonio H Lancha
Nov 18, 2021·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·André Luis Araujo Minari, Ronaldo V Thomatieli-Santos
Jan 20, 2022·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Bailey D PeckCharlotte A Peterson

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