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

Abstract

Immune dysregulation and inflammation by hepatic-resident leukocytes is considered a key step in disease progression of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis toward cirrhosis and hepatocellular carcinoma. Here, we provide a protocol for isolation and characterization of liver-resident immune cells from fine-needle biopsies obtained from a rodent model and humans. We describe steps for isolating leukocytes, cell sorting, and RNA extraction and sequencing. We then detail procedures for low-input mRNA sequencing analyses.

Keywords: Cell Isolation; Clinical Protocol; Flow Cytometry/Mass Cytometry; Health Sciences; Immunology; Molecular Biology; RNA-seq; Sequencing.