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Studying Macrophages in the Murine Steatotic Liver Using Flow Cytometry and Confocal Microscopy

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Tissue-Resident Macrophages

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2713))

Abstract

The study of macrophage functions in the context of metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction associated steatohepatitis (MASH) has been hampered by the fact that until recently all macrophages in the liver were thought to be Kupffer cells, the resident macrophages of the liver. With the advent of single-cell technologies, it is now clear that the steatotic liver harbors many distinct populations of macrophages, likely each with their own unique functions as well as subsets of monocytes and dendritic cells which can be difficult to discriminate from one another. Here, we detail the protocols we utilize to (i) induce MASLD/MASH in mice, (ii) isolate cells from the steatotic liver, and (iii) describe reliable gating strategies, which can be used to identify the different subsets of myeloid cells. Finally, we also discuss the issue of increased autofluorescence in the steatotic liver and the techniques we use to minimize this both for flow cytometry and confocal microscopy analyses.

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Acknowledgments

We thank Dr. Johnny Bonnardel who developed the in vivo perfusion protocol and the microscopy protocols on healthy liver tissue that were adapted here for use on the steatotic livers.

Author information

Authors and Affiliations

  1. Laboratory of Myeloid Cell Biology in Tissue Damage and Inflammation, VIB-UGent Center for Inflammation Research, Ghent, Belgium

    Zhuangzhuang Liu, Pieter A. Louwe & Charlotte L. Scott

  2. Department of Biomedical Molecular Biology, Faculty of Science, Ghent University, Ghent, Belgium

    Zhuangzhuang Liu, Pieter A. Louwe & Charlotte L. Scott

  3. Laboratory of Myeloid Cell Biology in Tissue Homeostasis and Regeneration, VIB-UGent Center for Inflammation Research, Ghent, Belgium

    Pieter A. Louwe

  4. Department of Chemical Sciences, Bernal Institute, University of Limerick, Castletroy, Co. Limerick, Ireland

    Charlotte L. Scott

Authors
  1. Zhuangzhuang Liu
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  2. Pieter A. Louwe
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  3. Charlotte L. Scott
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Corresponding author

Correspondence to Charlotte L. Scott .

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Editors and Affiliations

  1. Life & Medical Sciences Institute (LIMES), University of Bonn, Bonn, Germany

    Elvira Mass

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Liu, Z., Louwe, P.A., Scott, C.L. (2024). Studying Macrophages in the Murine Steatotic Liver Using Flow Cytometry and Confocal Microscopy. In: Mass, E. (eds) Tissue-Resident Macrophages. Methods in Molecular Biology, vol 2713. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3437-0_15

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  • DOI: https://doi.org/10.1007/978-1-0716-3437-0_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3436-3

  • Online ISBN: 978-1-0716-3437-0

  • eBook Packages: Springer Protocols

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