【摘要】 目的 制备低温冻存后转染EBV-LMP2、来源于人脐血的树突状细胞(dendritic cell，DC)疫苗，观察分析疫苗的形态和表型。方法 采用CD34 + 干细胞试剂盒，经MACS分选人脐血CD34 + 干细胞，GM-CSF和TNF-α诱导14天分化扩增为DC，低温冻存，再用包含EBV-LMP2基因的重组腺病毒感染DC制备EBV-LMP2-DC疫苗。倒置显微镜动态观察DC的形态，流式细胞仪检测EBV-LMP2-DC疫苗的CD86和CD83表型。结果 CD34 + 细胞在体外分化扩增过程中，数量逐渐增多，细胞形态由圆形逐渐变为不规则的毛刺状，并聚集成簇，发育为典型的成熟DC形态;冻存复温后，细胞回收率>85%;制备的EBV-LMP2-DC疫苗保持典型的成熟DC形态，细胞表型表达率CD86为71.69%，CD83为61.17%。结论 人脐血来源的DC，低温冻存复温后制备的EBV-LMP2-DC疫苗仍保持功能成熟DC形态和表型，为下一步应用EBV-LMP2-DC疫苗防治鼻咽癌的临床研究提供理想的疫苗和实验依据。

    关键词 树突状细胞 基因转染 表型 脐血 低温冻存

    The expression of CD86and CD83on EBV-LMP2dendritic cells vaccine derived from cord blood after cryopreservation

    Huang Yangliang，Mo Xuelian，Li Jie，et al.

    2000Grade Experimental Class Research Group，Shantou University Medical College，Shantou515041.

    【Abstract】 Objective To prepare human cord blood derived，cryopreserved dendrtic cells(DC)vaccine by thansfected with EBV-LMP2and study its morphology and phenotype.Methods CD34 + cells were purified from cord blood with CD34 + stem cell kit by mini MACS.The cells were induced into DC under the stimulation of GM-CSF and TNF-αfor14days and then cryopreserved in liquid nitrogen.EBV-LMP2-DC vaccine was prepared by transfected cryopreserved DC with recombined EBV-LMP2adenovirus.The morphology of DC vaccine was observed dynamically under the phase contrast microscope and the expression of CD86and CD83was detected with FACS.Reˉsults During the proliferation and differentiation ......