Gene expression profile at single-cell level of myeloid cells from the liver of mice after vertical sleeve gastrectomy (VSG)

We used droplet-based scRNA-seq to profile the gene expression of magnetically-sorted F4/80+ cells from the liver of Sham AL, Sham PF, and VSG mice. Two million F4/80+ cells from were individually labeled with cell multiplexing oligos (CMOs, 10X Genomics) to allow for the multiplexing of individual samples prior to pooling and gel bead-in-emulsion (GEM) preparation. Separate gene expression and CMO libraries were created and sequenced in a Novaseq S4 chip (2 x 150bp PE).

本研究采用基于液滴的单细胞RNA测序（droplet-based scRNA-seq）技术，对假手术自由进食（Sham AL）、假手术限食（Sham PF）及袖状胃切除术（VSG）小鼠的肝脏组织中磁分选得到的F4/80阳性细胞进行基因表达谱分析。我们从每份样本中分离获得200万个F4/80阳性细胞，分别使用细胞多重标记寡核苷酸（CMO，10X Genomics公司）进行标记，以实现在样本混合与乳液微滴包裹凝胶珠（GEM）制备前对单个样本进行多重标记。随后构建独立的基因表达文库与CMO文库，并在NovaSeq S4测序芯片上采用2×150bp双端测序模式完成测序。