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Related Experiment Video

Updated: Jun 12, 2025

Isolation of CD133+ Liver Stem Cells for Clonal Expansion
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Protocol for isolating CD163+ Kupffer cells from human liver resections.

Armando Andres Roca Suarez1, Marie-Laure Plissonnier1, Maud Michelet1

  • 1INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France.

STAR Protocols
|October 5, 2024
PubMed
Summary

This study details a method for isolating Kupffer cells (KCs), liver-resident macrophages, from human liver tissue. The protocol uses specific cell separation and identification techniques for accurate KC retrieval.

Keywords:
Cell BiologyCell isolationImmunology

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Area of Science:

  • Immunology
  • Cell Biology
  • Hepatology

Background:

  • The liver harbors diverse monocyte and macrophage populations.
  • Kupffer cells (KCs) are the primary resident macrophages in the liver, crucial for hepatic immunity and homeostasis.
  • Accurate isolation of KCs is essential for studying their functions and roles in liver diseases.

Purpose of the Study:

  • To provide a detailed protocol for the specific isolation of Kupffer cells from human liver resections.
  • To enable researchers to obtain pure KC populations for downstream analyses.
  • To facilitate the study of KC biology in the context of human liver health and disease.

Main Methods:

  • Human liver tissues are dissociated to single-cell suspensions.
  • Non-parenchymal cells are separated using a two-phase iodixanol density gradient.
  • Kupffer cells are isolated via positive selection based on CD163 expression.
  • Immunofluorescence is employed for procedure validation, detecting CD163 expression.

Main Results:

  • A reproducible protocol for isolating Kupffer cells from human liver tissue was established.
  • The method allows for the enrichment of CD163-positive Kupffer cells.
  • Validation confirms the specificity of the isolation procedure.

Conclusions:

  • This protocol provides a reliable method for obtaining Kupffer cells from human liver resections.
  • The isolated KCs can be used for various immunological and cellular studies.
  • This technique will aid in advancing the understanding of liver macrophage biology and associated pathologies.