An optimized protocol for isolation of hepatic leukocytes retrieved from murine and NASH liver biopsies

Immune dysregulation and inflammation by hepatic-resident leukocytes is considered a key step in disease progression of Non-alcoholic fatty liver disease (NAFLD) and Nonalcoholic steatohepatitis (NASH) toward cirrhosis and hepatocellular carcinoma (HCC). Here we provide a robust protocol for isolation and characterization of liver-resident immune cells from fine needle biopsies of rodent model and human. Various downstream applications can then be applied to gain an appreciation of the functional activity of liver-resident leukocyte populations. Overall design: Immune-cell FACS of liver needle biopsy tissue from mice using varying input cell numbers: 2,500 (n=3), 5,000 (n=3), 10,000 (n=3) and 20,000 (n=3) cells and bulk liver tissue as control (n=2)

肝脏驻留白细胞介导的免疫失调与炎症反应，被认为是非酒精性脂肪性肝病（Non-alcoholic fatty liver disease，NAFLD）与非酒精性脂肪性肝炎（Nonalcoholic steatohepatitis，NASH）向肝硬化及肝细胞癌（Hepatocellular carcinoma，HCC）进展过程中的关键环节。本研究提供了一套稳健的实验方案，可从啮齿类动物模型及人类的肝脏细针活检组织中分离并鉴定肝脏驻留免疫细胞。后续可开展多种下游实验，以解析肝脏驻留白细胞群体的功能活性。

整体实验设计：针对小鼠肝脏针吸活检组织，采用不同输入细胞数开展免疫细胞荧光激活细胞分选（Fluorescence Activated Cell Sorting，FACS），分组为2500个细胞（n=3）、5000个细胞（n=3）、10000个细胞（n=3）及20000个细胞（n=3），并以全肝组织作为对照（n=2）。