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Liver Macrophage Isolation by Flow Cytometry Sorting

Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 2164)

Abstract

Liver macrophages are immune cells that under steady state conditions play important roles in maintaining tissue homeostasis. But these cells are also intricately linked to the pathogenesis of obesity-related diseases such as nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), and type 2 diabetes (T2D). Described herein is a protocol to isolate liver macrophages by FACS for downstream molecular analysis. Such studies of macrophages are important for elucidating their role in the pathogenesis of these metabolic diseases.

Key words

FACS Isolation Insulin sensitivity Macrophages Liver Obesity Diabetes Inflammation Innate immunity 

Notes

Acknowledgments

Valerio Azzimato and Emelie Barreby are acknowledged for providing cells.

References

  1. 1.
    Schulz C, Gomez Perdiguero E, Chorro L, Szabo-Rogers H, Cagnard N, Kierdorf K, Prinz M, Wu B, Jacobsen SE, Pollard JW, Frampton J, Liu KJ, Geissmann F (2012) A lineage of myeloid cells independent of Myb and hematopoietic stem cells. Science 336(6077):86–90.  https://doi.org/10.1126/science.1219179CrossRefPubMedGoogle Scholar
  2. 2.
    Scott CL, Zheng F, De Baetselier P, Martens L, Saeys Y, De Prijck S, Lippens S, Abels C, Schoonooghe S, Raes G, Devoogdt N, Lambrecht BN, Beschin A, Guilliams M (2016) Bone marrow-derived monocytes give rise to self-renewing and fully differentiated Kupffer cells. Nat Commun 7:10321.  https://doi.org/10.1038/ncomms10321CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kondo H, Saito K, Grasso JP, Aisen P (1988) Iron metabolism in the erythrophagocytosing Kupffer cell. Hepatology 8(1):32–38CrossRefGoogle Scholar
  4. 4.
    Wong CH, Jenne CN, Petri B, Chrobok NL, Kubes P (2013) Nucleation of platelets with blood-borne pathogens on Kupffer cells precedes other innate immunity and contributes to bacterial clearance. Nat Immunol 14(8):785–792.  https://doi.org/10.1038/ni.2631CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Skogh T, Blomhoff R, Eskild W, Berg T (1985) Hepatic uptake of circulating IgG immune complexes. Immunology 55(4):585–594PubMedPubMedCentralGoogle Scholar
  6. 6.
    Scott CL, Guilliams M (2018) The role of Kupffer cells in hepatic iron and lipid metabolism. J Hepatol 69(5):1197–1199.  https://doi.org/10.1016/j.jhep.2018.02.013CrossRefPubMedGoogle Scholar
  7. 7.
    Kazankov K, Jorgensen SMD, Thomsen KL, Moller HJ, Vilstrup H, George J, Schuppan D, Gronbaek H (2019) The role of macrophages in nonalcoholic fatty liver disease and nonalcoholic steatohepatitis. Nat Rev Gastroenterol Hepatol 16(3):145–159.  https://doi.org/10.1038/s41575-018-0082-xCrossRefPubMedGoogle Scholar
  8. 8.
    Gadd VL, Skoien R, Powell EE, Fagan KJ, Winterford C, Horsfall L, Irvine K, Clouston AD (2014) The portal inflammatory infiltrate and ductular reaction in human nonalcoholic fatty liver disease. Hepatology 59(4):1393–1405.  https://doi.org/10.1002/hep.26937CrossRefPubMedGoogle Scholar
  9. 9.
    Aparicio-Vergara M, Tencerova M, Morgantini C, Barreby E, Aouadi M (2017) Isolation of Kupffer cells and hepatocytes from a single mouse liver. Methods Mol Biol 1639:161–171.  https://doi.org/10.1007/978-1-4939-7163-3_16CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Medicine, Integrated Cardio Metabolic CenterKarolinska InstitutetHuddingeSweden

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