Mater Research Institute-The University of Queensland, Translational Research Institute, Woolloongabba, QLD 4102, Australia.
Myeloma Research Laboratory, Adelaide Medical School, Faculty of Health and Medical Sciences, University of Adelaide, North Terrace, Adelaide, SA 5005, Australia; South Australian Health and Medical Research Institute, PO Box 11060, Adelaide, SA 5001, Australia.
Cell Rep. 2021 Nov 23;37(8):110058. doi: 10.1016/j.celrep.2021.110058.
Mouse hematopoietic tissues contain abundant tissue-resident macrophages that support immunity, hematopoiesis, and bone homeostasis. A systematic strategy to characterize macrophage subsets in mouse bone marrow (BM), spleen, and lymph node unexpectedly reveals that macrophage surface marker staining emanates from membrane-bound subcellular remnants associated with unrelated cells. Intact macrophages are not present within these cell preparations. The macrophage remnant binding profile reflects interactions between macrophages and other cell types in vivo. Depletion of CD169 macrophages in vivo eliminates F4/80 remnant attachment. Remnant-restricted macrophage-specific membrane markers, cytoplasmic fluorescent reporters, and mRNA are all detected in non-macrophage cells including isolated stem and progenitor cells. Analysis of RNA sequencing (RNA-seq) data, including publicly available datasets, indicates that macrophage fragmentation is a general phenomenon that confounds bulk and single-cell analysis of disaggregated hematopoietic tissues. Hematopoietic tissue macrophage fragmentation undermines the accuracy of macrophage ex vivo molecular profiling and creates opportunity for misattribution of macrophage-expressed genes to non-macrophage cells.
小鼠造血组织中含有丰富的组织驻留巨噬细胞，这些细胞支持免疫、造血和骨稳态。一项系统的策略来描述小鼠骨髓（BM）、脾脏和淋巴结中的巨噬细胞亚群，出人意料地揭示了巨噬细胞表面标记染色源自与无关细胞相关的膜结合亚细胞残余物。在这些细胞制剂中不存在完整的巨噬细胞。巨噬细胞残余结合谱反映了体内巨噬细胞与其他细胞类型之间的相互作用。体内耗尽 CD169 巨噬细胞会消除 F4/80 残余物的附着。残余物限制的巨噬细胞特异性膜标记物、细胞质荧光报告物和 mRNA 都在非巨噬细胞中检测到，包括分离的干细胞和祖细胞。对 RNA 测序（RNA-seq）数据的分析，包括公开可用的数据集，表明巨噬细胞碎片化是一种普遍现象，会混淆分散造血组织的批量和单细胞分析。造血组织中巨噬细胞的碎片化破坏了巨噬细胞体外分子分析的准确性，并为巨噬细胞表达的基因错误归因于非巨噬细胞创造了机会。
