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. Overall design: 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.

非酒精性脂肪性肝炎（Non-Alcoholic Steatohepatitis, NASH）与肝脏巨噬细胞组成的显著改变密切相关。为深入解析这一细胞异质性，我们对喂食16周高脂高糖饮食的小鼠肝脏髓系细胞开展了单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）。
实验整体设计：选取3只C57CL/6雄性小鼠，喂食高脂高糖饲料（Research Diets公司，货号D12492）16周；随后通过门静脉灌注及percoll密度梯度离心法从肝脏中分离免疫细胞，对活单细胞进行流式分选，通过门控策略筛选CD11bhi、F4/80int与CD11bint、F4/80hi两个细胞群，并借助Ly6G与siglecF标记排除嗜酸性粒细胞及中性粒细胞。将所得巨噬细胞样品按1:1比例混合，合并3只小鼠的样品以制备单个scRNA-seq测序样本。