Myeloid immune cells, e.g. macrophages, play a crucial role in the progression of non-alcoholic fatty liver disease (NAFLD), and thus significantly contribute to the development of non-alcoholic steatohepatitis (NASH). However, due to its complex phenotype and high heterogeneity, this cell type is particularly difficult to isolate and analyse. Single cell RNA sequencing (scRNA-seq) is a potential solution to this problem as it is able to characterise diverse hepatic cell types and compare gene expression profiles of both NASH and non-NASH samples.
The aim of this study was to isolate and analyse myeloid cell populations (specifically macrophages) in a mouse model using microdroplet-based scRNA-seq.
A single cell solution of myeloid leukocytes was extracted from mouse liver by gradient centrifugation and subsequent fluorescence-activated cell sorting. After this preparation, a barcoded single cell library was generated and followed by sequencing on an Illumina NextSeq 500. Further bioinformatic analysis was then performed on the sequence generated.
This analysis found that the livers of NASH mice possessed a significantly higher number of myeloid-derived cell populations. These populations also adopted an inflammatory phenotype characteristic of the innate immune dysregulation associated with the progression of NAFLD into NASH. This finding highlights the usefulness of scRNA-seq in improving clinical understanding of NASH development.