Single Cell RNA Sequencing in NASH

Methods Mol Biol. 2022:2455:181-202. doi: 10.1007/978-1-0716-2128-8_15.

Abstract

Single cell RNA sequencing (scRNA-seq) allows to uncover cellular heterogeneity and the identification of novel subpopulations. In non-alcoholic steatohepatitis (NASH), scRNA-seq is particularly powerful to understand non-parenchymal cell heterogeneity in the liver, e.g. for inflammatory cells. Myeloid immune cells, particularly macrophages, play a critical role in response of the innate immune system and significantly contribute to the progression of fatty liver disease. Due to their high heterogeneity and complex phenotypes, their functional role in health and disease is difficult to analyze. Here, we describe the isolation and analysis of myeloid cell populations from mouse liver using microdroplet-based scRNA-seq. This approach allows the identification and characterization of different hepatic cell types, exemplified here by hepatic macrophage populations, as well as analyses of differentially expressed genes between samples (e.g., cells from healthy or NASH livers).

Keywords: 10× Genomics; Fibrosis; Macrophages; Myeloid immune cells; NAFLD; NASH; Single cell RNA sequencing (scRNA-seq); Transcriptomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Kupffer Cells / metabolism
  • Liver / metabolism
  • Macrophages / metabolism
  • Mice
  • Non-alcoholic Fatty Liver Disease* / genetics
  • Non-alcoholic Fatty Liver Disease* / metabolism
  • Sequence Analysis, RNA