Transcriptome of human primary macrophages from healthy controls after long-term myelin phagocytosis. Transcriptome of human primary macrophages from healthy controls after long-term myelin phagocytosis

CD14+ monocytes were isolated from the blood of healthy human donors by Ficoll density-gradient centrifugation, magnetic cell sorting and LS column systems. CD14+ monocytes were in vitro differentiated to macrophages by culturing the cells for 7 days in RPMI medium containing 10% FCS, 1% penicillin/streptomycin, 1% glutamine, 1% Fungizone and supplemented with 50 ng/ml human macrophage-colony stimulating factor (M-CSF) in the presence or absence of myelin isolated from brain tissue of 6-8 week-old wild-type C57BL6/J mice. Overall design: Comparative gene expression profiling analysis of human monocyte-derived macrophages differentiated in the absence or presence of myelin from brain tissue of wild-type C57BL6/J mice

CD14+单核细胞（CD14+ monocytes）通过Ficoll密度梯度离心（Ficoll density-gradient centrifugation）、磁细胞分选（magnetic cell sorting）及LS柱系统（LS column systems）从健康人类供者外周血中分离获得。将CD14+单核细胞体外诱导分化为巨噬细胞：将细胞培养于含10%胎牛血清（FCS）、1%青霉素/链霉素、1%谷氨酰胺、1%Fungizone的RPMI培养基中，持续培养7天，同时添加50 ng/ml 人巨噬细胞集落刺激因子（human macrophage-colony stimulating factor, M-CSF），并分别设置加入或不加入从6-8周龄野生型C57BL6/J小鼠脑组织中分离的髓鞘（myelin）的培养条件。实验整体设计：对分别在添加或不添加野生型C57BL6/J小鼠脑组织来源髓鞘的条件下分化得到的人单核细胞源性巨噬细胞进行比较基因表达谱分析。