Isolation of non-parenchymal cells from the mouse liver

Isaac Mohar, Katherine J. Brempelis, Sara A. Murray, Mohammad Ebrahimkhani, I. Nicholas Crispe

Research output: Chapter in Book/Report/Conference proceedingChapter

19 Scopus citations

Abstract

Hepatocytes comprise the majority of liver mass and cell number. However, in order to understand liver biology, the non-parenchymal cells (NPCs) must be considered. Herein, a relatively rapid and efficient method for isolating liver NPCs from a mouse is described. Using this method, liver sinusoidal endothelial cells, Kupffer cells, natural killer (NK) and NK-T cells, dendritic cells, CD4+ and CD8+ T cells, and quiescent hepatic stellate cells can be purified. This protocol permits the collection of peripheral blood, intact liver tissue, and hepatocytes, in addition to NPCs. In situ perfusion via the portal vein leads to efficient liver digestion. NPCs are enriched from the resulting single-cell suspension by differential and gradient centrifugation. The NPCs can by analyzed or sorted into highly enriched populations using flow cytometry. The isolated cells are suitable for flow cytometry, protein, and mRNA analyses as well as primary culture.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages3-17
Number of pages15
Volume1325
DOIs
StatePublished - 2015
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1325
ISSN (Print)10643745

Keywords

  • Cell isolation
  • Hepatic stellate cells
  • Kupffer cells
  • Liver
  • Perfusion
  • Sinusoidal endothelial cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

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  • Cite this

    Mohar, I., Brempelis, K. J., Murray, S. A., Ebrahimkhani, M., & Crispe, I. N. (2015). Isolation of non-parenchymal cells from the mouse liver. In Methods in Molecular Biology (Vol. 1325, pp. 3-17). (Methods in Molecular Biology; Vol. 1325). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-2815-6_1