人外周血树突状细胞体外诱导扩增的研究  

Experimental Study on Induction and Expansion of Dendritic Cells from Human Peripheral Blood In Vitro

作　　者：吴楠 刘薇[2] 卢光琇[2] 

机构地区：[1]深圳南山区疾病预防控制中心,广东深圳518054 [2]中南大学干细胞研究所,湖南长沙410078

出　　处：《现代生物医学进展》2012年第25期4948-4949,4997,共3页Progress in Modern Biomedicine

摘　　要：目的:探讨可用于临床治疗功能成熟的DC体外扩增的优化培养方案。方法:胎牛血清培养基联合细胞因子rhGM-CSF(100ng/mL)和rhIL-4(50 ng/mL)扩增人外周血分离的单个核细胞,细胞培养分别按5×106/mL、6×106/mL和7×106/mL的密度,加入6孔培养板。第6d加入rhTNF-a(100 ng/mL)联合培养,分别于第6 d,第9 d和12 d收获细胞。从形态学、细胞表面标志方面进行鉴定。结果:显微镜观察,经过9 d诱导后,培养细胞具有典型树突细胞外形。流式细胞仪分析,6×106/mL密度的细胞培养组培养到第9天最宜。结论:细胞具有典型的DC的形态特征,细胞表型及功能实验证实其DC的特性,说明建立的血清培养基联合细胞因子rhGM-CSF、rhIL-4和rhTNF-a体外诱导DC的方法是切实可行的。Objective： To investigate a culture system for DC expansion.Methods： Using serum medium with rhGM-CSF（100ng/mL） and rhIL-4（50ng/mL） for 6 days,rhTNF-a（100ng/mL） was added on the 6th day of culture.Human peripheral blood were seeding at the concentration of 5×106/mL,6×106/mL and 7×106/mL in 6-well flat-bottomed plates.Suspension cells were harvested at days 6,9 and 12 and analysed on morphology,phenotype observed.Result： Cells on day 9 of culture displayed DC-like morphology.Phenotype analysis on DCs acquired showed that the cells of 6×106/mL cultured for 9 days contained a higher percentage of DC cells.Conclusion： The results suggested the culture method was feasible.

关 键 词：树突状细胞 细胞培养 细胞因子 

分 类 号：R392.12[医药卫生—免疫学] Q813[医药卫生—基础医学]