TY - CHAP
T1 - Isolation of non-parenchymal cells from the mouse liver
AU - Mohar, Isaac
AU - Brempelis, Katherine J.
AU - Murray, Sara A.
AU - Ebrahimkhani, Mohammad R.
AU - Crispe, I. Nicholas
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2015.
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
KW - Cell isolation
KW - Hepatic stellate cells
KW - Kupffer cells
KW - Liver
KW - Perfusion
KW - Sinusoidal endothelial cells
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U2 - 10.1007/978-1-4939-2815-6_1
DO - 10.1007/978-1-4939-2815-6_1
M3 - Chapter
C2 - 26450375
AN - SCOPUS:84943743994
T3 - Methods in Molecular Biology
SP - 3
EP - 17
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -