title: Isolation of Non-parenchymal Cells from the Mouse Liver.
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.
authors.affiliation: Gradient, 600 Stewart St., Suite 1900, Seattle, WA, 98101, USA. imohar@gradientcorp.com.
authors.forename: Isaac
authors.initials: I
authors.lastname: Mohar
authors.affiliation: Department of Global Health, University of Washington, Seattle, WA, USA.
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Brempelis
authors.affiliation: Systems Immunology, Benaroya Research Institute, Seattle, WA, USA.
authors.forename: Sara A
authors.initials: SA
authors.lastname: Murray
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: Mohammad R
authors.initials: MR
authors.lastname: Ebrahimkhani
authors.affiliation: Department of Pathology, University of Washington, Seattle, WA, USA.
authors.forename: I Nicholas
authors.initials: IN
authors.lastname: Crispe
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology; D042783:Endothelial Cells / Q000166:cytology; D007694:Killer Cells, Natural / Q000166:cytology; D007728:Kupffer Cells / Q000166:cytology; D008099:Liver / Q000166:cytology*; D051379:Mice; D008967:Molecular Biology / Q000379:methods*
keywords: Cell isolation; Hepatic stellate cells; Kupffer cells; Liver; Perfusion; Sinusoidal endothelial cells