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.

Intestinal Alkaline Phosphatase Attenuates Alcohol-Induced Hepatosteatosis in Mice

“…Our findings suggest that oral supplementation of IAP can prevent liver injury due to alcohol intake in mice. Alcohol induced liver cell damage results in elevated levels of serum ALT levels in human and rodents [32] and in clinical settings ALT levels are proportional to the degree of liver injury [33]. Both pre- and simultaneous IAP treatment with alcohol significantly lowered the serum ALT levels, while giving the IAP after the alcohol failed to block the elevations in ALT.…”

“…Hepatocytes were isolated from control C57BL/6 mice and hepatic stellate cells isolated from control and ethanol +/− IAP-treated mice by enzymatic digestion and Percoll density gradient centrifugation with modifications [32]. The portal vein was perfused in situ with 30 mL of HBSS (without Ca2+ and Mg2+) and 30 mL of 0.05% collagenase B (Roche Diagnostics, Indianapolis, IN), respectively, at 37 °C with a flow rate of 6 ml/min.…”

Cachexia and fibrosis are costs of chronic IL-1R-mediated disease tolerance inT. gondiiinfection

“…Preparation of tissues for flow cytometric analysis was based on previously published protocols. (Mohar et al, 2015) Briefly, following euthanasia by CO2 and cardiac exsanguination at 9 weeks post-infection, mice were perfused through the heart with 10 mL perfusion buffer (HBSS, 5 mM HEPES, 0.5 mM EDTA), and liver, epigonadal visceral white adipose tissue, and quadriceps were excised. Following mechanical disruption of the tissue, tissue was collagenase-digested for 30 minutes at 37°C (shaking at 130 RPM) and filtered.…”

Immune function of nonparenchymal liver cells

“…The liver comprises parenchymal and non-parenchymal cells, which account for 80 and 20% of the total liver cells, respectively. The non-parenchymal cells mainly include liver sinusoidal endothelial cells (LSECs), dendritic cells (DCs), hepatic stellate cells (HSCs), and Kupffer cells (KCs) (Mohar et al, 2015). These cells have different structural functions and sources of differentiation, and they regulate the local and systemic immune function.…”