脐血源树突状细胞的扩增及其杀肿瘤细胞效应研究

Studies In Vitro on Proliferating Dendritic Cell Derived from Cord Blood and Its Role in Antitumor Immunity

目的:探讨不同细胞因子组合对脐带血来源的树突状细胞(Dendritic cells,DC)扩增率的影响,优化扩增方案,并对所诱导的脐带血DC的生物学特征进行鉴定。方法:收集健康孕妇正常顺产儿脐带血,用淋巴细胞分离液分离单个核细胞,加入不同组合的细胞因子,联合诱导脐带血单个核细胞前体细胞分化为DC。然后对诱导细胞进行流式细胞仪检测,其中包括CD83,CD1a在DC表面的表达。激光共聚焦显微镜观察DC的形态。用MTT法检测所诱导的DC活化的T细胞对肿瘤细胞的杀伤效应。结果:单个核细胞经细胞因子诱导后第3d成均匀散在分布的细胞,部分细胞变形,胞体拉长;第7d可见胞浆突起,细胞形态不规则,呈疏松贴壁生长或悬浮生长;第12 d细胞集落样生长,具有典型的DC特征。检测所诱导的DC活化的T细胞对肿瘤细胞的杀伤效应,实验组DC活化的T淋巴细胞对肿瘤细胞的杀伤效应明显优于对照组。结论:采取细胞因子GM-CSF、IL-4、FL、SCF联合刺激成功诱导出形态典型、纯度较高的脐带血DC,收获的细胞数比常规方案(加细胞因子GM-CSF、IL-4)明显增多。优化方案所诱导的DC能增强T淋巴细胞对肿瘤细胞的杀伤效应。

Objective To investigate the way of inducing dendritic cell(DC) from precursor in cord blood by using cytokines and role of the DC in antitumor immunity.Methods Mononuclear cells derived from human umbilical cord blood were separated,and the progenitor cells of DC were induced into DC by adding different combinations of cytokines.CD83 and CD1a expression of DC surface were detected by using Flow cytometer Morphology of DC was detected by using Laser Confocal Microscopy.MTT assay was used to detect the killing effect on tumor cells of T cell which were activated by the induced DC.Results Dendritic cells induced from cord blood cells had typical morphological properties of DC.The killing effect on tumor cells by T cells activated by the induced DC of the experimental group was significantly better than control groups.Concclusion DC induced by cytokines jointly with GM-CSF,IL-4,FL,and SCF,superior to the regular programme(with cytokine GM-CSF,IL-4). The DC can enhance T cell killing effect on tumor cells.