Isolation of non-parenchymal cells from the mouse liver

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

Research output: Chapter in Book/Report/Conference proceedingChapter

44 Scopus citations


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.
Number of pages15
StatePublished - 2015

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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