Transcriptomic profile of dermal and hypodermal CCR2+ macrophages, CCR2- macrophages and dendritic cells from C57BL/6 mice

In order to explore and compare the transcriptomic profile of CCR2+ macrophages, CCR2- macrophages and dendritic cells present in the dermis and hypodermis of C57BL/6 mice, isolated cells were subjected to RNA-sequencing analysis. Back skins from C57BL/6 female mice were harvested and the dermal and hypodermal layers isolated by mechanical force and enzymatic treatments. Cells were liberated from each skin layers after enzymatic digestion of the tissues. Macrophages (CD45+ CD11b+ EpCAM-Ly6C-lo CD11c-lo/mid CD64+) expressing or not CCR2 as well as dendritic cells (CD45+ CD11b+ EpCAM-Ly6C-lo CD11c-lo/mid CD64- MHC-II+) were isolated by fluorescence-activated cell sorting (FACS) and their transcriptome assessed by RNA-sequencing.为探索并比较C57BL/6小鼠真皮及皮下组织中CCR2阳性巨噬细胞、CCR2阴性巨噬细胞与树突状细胞的转录组特征，研究人员对分离得到的细胞开展了RNA测序（RNA-sequencing）分析。研究人员获取C57BL/6雌性小鼠的背部皮肤，通过机械剥离与酶消化法分离真皮层与皮下组织层；对各皮肤层的组织进行酶解后，释放出其中的细胞。随后通过荧光激活细胞分选术（fluorescence-activated cell sorting, FACS）分离得到两类巨噬细胞：表达CCR2的巨噬细胞（CD45+ CD11b+ EpCAM-Ly6C-lo CD11c-lo/mid CD64+）与不表达CCR2的巨噬细胞，以及树突状细胞（CD45+ CD11b+ EpCAM-Ly6C-lo CD11c-lo/mid CD64- MHC-II+），并通过RNA测序对其转录组进行检测分析。