Kassianos Andrew J, Jongbloed Sarah L, Hart Derek N J, Radford Kristen J
Mater Medical Research Institute, South Brisbane, QLD, Australia.
Methods Mol Biol. 2010;595:45-54. doi: 10.1007/978-1-60761-421-0_3.
Human blood dendritic cells (DCs) are a rare, heterogeneous cell population that comprise approximately 1% of circulating peripheral blood mononuclear cells (PBMCs). Their isolation has been confounded by their scarcity and lack of distinguishing markers and their characterisation perplexed by the recent discovery of phenotypic and functionally distinct subsets. Human blood DCs are broadly defined as leukocytes that are HLA-DR positive and lack expression of markers specific for T cell, B cell, NK cell, monocyte and granulocyte lineages. They can be subdivided into the CD11c(-) (CD123(+)CD303(+)CD304(+)) plasmacytoid DC and CD11c(+) myeloid DC, which can be further subdivided into three subsets based on differential expression of CD1c, CD141 and CD16. DC can be isolated from peripheral blood by using an initial density gradient centrifugation step to enrich for mononuclear cells followed by immunomagnetic depletion of cells expressing markers specific for leukocyte lineages and undesired DC subsets. Subsequent flow cytometry-based cell sorting allows the isolation of highly pure individual DC subsets that can then be used for functional studies.
人血树突状细胞（DCs）是一种罕见的异质性细胞群体，约占循环外周血单核细胞（PBMCs）的1%。它们的稀缺性、缺乏特异性标志物以及最近发现的表型和功能不同的亚群，使得其分离和鉴定都面临困难。人血DCs被广泛定义为HLA-DR阳性且缺乏T细胞、B细胞、NK细胞、单核细胞和粒细胞谱系特异性标志物表达的白细胞。它们可细分为CD11c(-)（CD123(+)CD303(+)CD304(+)）浆细胞样DC和CD11c(+)髓样DC，后者可根据CD1c、CD141和CD16的差异表达进一步细分为三个亚群。通过初始密度梯度离心步骤富集单核细胞，然后对表达白细胞谱系特异性标志物的细胞和不需要的DC亚群进行免疫磁珠去除，可从外周血中分离出DC。随后基于流式细胞术的细胞分选可分离出高度纯化的单个DC亚群，然后用于功能研究。
