Picturing Polarized Myeloid Phagocytes and Regulatory Cells by Mass Cytometry.
Picturing Polarized Myeloid Phagocytes and Regulatory Cells by Mass Cytometry.
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通过质谱流式细胞仪描绘极化的骨髓吞噬细胞和调节细胞。
DOI：
10.1007/978-1-4939-9454-0_14
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Irish,JonathanM
中科院分区：
文献类型：
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作者：
Roussel,Mikael;Bartkowiak,Todd;Irish,JonathanM
免疫单核细胞/吞噬细胞系统（MPS）包括骨髓谱系的许多细胞亚群，包括单核细胞、巨噬细胞和树突状细胞（DC）群体，其在表型和功能上都是异质的。以前，我们用多参数质谱细胞术（CyTOF）表征了这些不同的MPS表型。为了广泛表征单核细胞、巨噬细胞和树突状细胞，设计了CyTOF面板以测量35种身份、活化和极化标志物。在这里，我们提供了一个协议，以定义一个参考地图的骨髓室，包括样品制备，产生参考细胞亚群的单核细胞/吞噬细胞系统。特别地，我们关注单核细胞衍生的巨噬细胞，其在体外用细胞因子刺激进一步极化（即，M-CSF、GM-CSF、IL-4、IL-10、IFNγ和LPS），以及单核细胞来源的DC和骨髓来源的抑制细胞（MDSC），其在体外从人骨髓和/或外周血产生。
The immune monocyte/phagocyte system (MPS) includes numerous cell subsets of the myeloid lineage including monocyte, macrophage, and dendritic cell (DC) populations that are heterogeneous both phenotypically and functionally. Previously, we characterized these diverse MPS phenotypes with multi-parametric mass cytometry (CyTOF). In order to expansively characterize monocytes, macrophages, and dendritic cells, a CyTOF panel was designed to measure 35 identity-, activation-, and polarization-markers. Here we provide a protocol to define a reference map for the myeloid compartment, including sample preparation, to produce reference cell subsets from the monocyte/phagocyte system. In particular, we focused on monocyte-derived macrophages that were further polarized in vitro with cytokine stimulation (i.e., M-CSF, GM-CSF, IL-4, IL-10, IFNγ, and LPS), as well as monocyte-derived DCs, and myeloid-derived suppressor cells (MDSCs), generated in vitro from human bone marrow and/or peripheral blood.