Dynamic Shifts in the Composition of Resident and Recruited Macrophages Influence Tissue Remodeling in NASH

NASH is associated with signficant changes in liver macrophage composition. To gain insight into this heterogenity we performed scRNA on liver myeloid cells from mice fed a HFD for 16 weeks. Three C57CL/6 male mice were fed High fat, high sucrose diet (Research Diets -cat# D12492) for 16 weeks after which immune cells were isolated from the liver using portal vein perfusion and a percoll gradient protocol. The cells underwent flow sorting on live singlet cells and were gated on CD11bhi, F4/80int and CD11bint, F4/80hi populations. Eosinophils and neutrophils were removed using Ly6G and siglecF exlusion. Samples of these macrophages were mixed 1:1 and the 3 mouse samples were pooled to create a signal sample for scRNA seq.