脐血CD34^+细胞体外诱导分化为树突状细胞及其扩增  

Induction and amplification of dendritic cells derived from cord blood CD34^+ cellsin vitro

作　　者：陈雪华[1] 黎皓[1] 俞焙秦[1] 郑重[1] 柯山[1] 朱正纲[1] 顾琴龙[1] 刘炳亚[1] 

机构地区：[1]上海交通大学医学院瑞金医院外科上海消化外科研究所,上海200025

出　　处：《上海交通大学学报（医学版）》2007年第5期516-519,共4页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30170915;30570828;30670939)~~

摘　　要：目的探索脐血CD34+细胞体外诱导分化为树突状细胞及其扩增、培养大量树突状细胞(DCs)的方法。方法Ficoll分离脐血单个核细胞,免疫微磁珠法分离纯化CD34+细胞,以rhIL-4、GM-CSF、rhSCF、rhFlt-3Ligand、TGF-β1及TNF-α体外诱导培养14d,进行形态学观察、表型检测及上清液IL-12测定。结果体外培养14d后,细胞呈现典型的成熟DCs形态学及表型特征:胞体表面粗糙,可见大量突起,高水平表达CD1a、CD11c、CD40、CD80、CD83、CD86和MHCⅡ类分子;同时,IL-12浓度随着培养时间的增加而升高,且在加入TNF-α刺激成熟后,IL-12浓度较未成熟时显著升高(P<0.05)。结论体外联合运用rhIL-4、GM-CSF、rhSCF、rhFlt-3Ligand、TGF-β1、TNF-α,能够成功诱导脐血CD34+细胞分化为大量活力好的DCs。Objective To investigate the induction and amplification of dendritic cells derived from cord blood CD34 ^＋ cells in vitro and establish an effective culture system for dendritic cells. Methods Core blood mononuclear cells were isolated by Ficoll. CD34 ^＋ cells were purified from mononuclear cells by MACS and then cultured in a complete medium containing rhIL-4, GM-CSF, rhSCF, rhFh-3 Lig- and, TGF-β1 and TNF-α. After 14 d of incubation, the morphology of the cells was observed, the phonotype was examined and the IL-12 level in the medium was measured. Results DCs exhibited typical morphology and phonotype after 14 d. The surface of the cell body was rough with a large number of prominence, and CDla, CD11c, CD40, CD80, CD83, CD86 and MHC Ⅱ molecule were in high-level expression. Meanwhile, the level of IL-12 rised with the time of culture, and was significantly increased in the cell mature state after culture with TNF-α compared with that in the cell immature state（P〈0.05）. Conclusion The in vitro culture system with a combination of cytokines, such as rhIL-4, GM-CSF, rhSCF, rhFlt-3Ligand, TGF-β1 and TNF-α can effectively induce cord blood CD34 ^＋ cells to differentiate into dendritic cells.

关 键 词：树突状细胞 脐血 CD34 诱导 扩增 

分 类 号：R392[医药卫生—免疫学]