摘要
以人浓缩白细胞来源的CD14+单核细胞为前体,建立体外快速培养树突状细胞(dendritic cell,DC)的方法。采用密度梯度离心和MACS磁珠分选系统,收集高纯度的CD14+单核细胞;以rGM-CSF、rIL-4联合分化2天诱导不成熟DC,再将分化后的细胞以rTNF-α、IL-1β、IL-6、PGE2共同活化2天得到成熟DC。流式细胞仪检测结果表明,分化2天的不成熟DC具有吞噬能力,且表型HLA-DR、CD40、CD80表达在80%以上,CD83、CD86基本不表达,成熟后的DC能够激活T细胞增殖,HLA-DR表达增高,CD83、CD86表达占85%。
A ＂2 ＋2 days＂ fast protocol for the generation of dendritic cells（DC ） from high purity human monocytes in vitro has been established. During the 2-step differentiation and activation process, we demonstrated that 2 days of culture with GM-CSF and IL-4 were sufficient to generate immature DCs capable of antigen uptake. Similarly the mature DCs were activated with a cocktail of rTNF-ct, IL-113, IL-6, and PGE2 from immature DC in 2 days. This ＂2＋2＂ fast DC had the same phenotype and function as the ＂6 ＋ 2＂ standard DC.
基金
国家自然科学基金面上项目(30771951)
中国科学院"百人计划"项目
中国科学院知识创新工程重要方向项目(KSCX1-YW-R-16)
