脐静脉内皮细胞诱导脐血单个核细胞向树突状细胞的分化  

Human umbilical blood mononuclear cells differentiated into dendritic cells by induction of umbilical vein endothelial cells

作　　者：齐莹莹 杜英[2] 李倩如[2] 陈惠萍[3] 徐虹[4] 周云[5] 董子明[2] 

机构地区：[1]郑州市人民医院检验科,450000 [2]郑州大学基础医学院微生物学与免疫学教研室,450001 [3]郑州大学基础医学院病理生理学教研室,450001 [4]河南省肿瘤研究所,郑州450000 [5]河南省人民医院肿瘤科,郑州450000

出　　处：《重庆医学》2012年第4期355-356,359,F0003,共4页Chongqing medicine

摘　　要：目的探讨人脐静脉内皮细胞对人脐血单个核细胞(UBMCs)向树突状细胞(DC)分化的影响,建立更便捷、更经济体外培养DC的方法。方法胶原酶消化分离出脐静脉内皮细胞(HUVEC),经鉴定和体外培养扩增后,使其平铺生长于Tran-swell板上室底部,将UBMCs接种到Transwell小室内,同时加入EC9706细胞冻融形成的蛋白质抗原,与HUVEC共培养,使UBMCs与HUVEC紧密接触并从Transwell板上室迁移到下室,收集Transwell板下室细胞,流式细胞术检测下室细胞DC相关表面标志CD80、CD40、CD83、CD1a和MHC分子;通过MTT法检测肿瘤负载的UBDC诱导CTL对靶细胞的特异性杀伤作用。结果经HUVEC诱导并负载了肿瘤细胞抗原的DC CD80、CD40、CD83、CD1a和MHC-Ⅱ类分子表达增加,产生的细胞毒性T细胞(CTL)可特异性杀伤食管癌细胞EC9706;与细胞因子诱导DC活化的CTL对EC9706的杀伤率比较差异无统计学意义(P>0.05)。结论利用Transwell小室以人原代脐静脉内皮细胞诱导脐血单个核细胞分化的DC是可行的一种实验方法。Objective To investigate the influence of human umbilical vein endothelial cells（HUVEC） on the differentiation of umbilical blood mononuclear cells（UBMCs） to dendritic cells（DCs） for establishing the more convenient,faster and more economic in vitro culture method of DCs.Methods HUVEC was digested and isolated by collagenase.After identification and in vitro culture and amplification,HUVEC were tiled and grown in the bottom of Transwell plate.UBMCs were inoculated to Transwell small indoor,and at the same time protein antigen formed by EC9706 cell freezing and thawing was added and cocultured with HUVEC.UBMCs were closely contacted with HUVEC and migrated from Transwell upper chamber to the below chamber.The chamber cells were collected under Transwell plate.The dendritic cells related surface marker CD83,CD80,CD40,CD1a and MHC molecules were detected by the flow cytometry.The MTT assay was adopted to detect the specific target cell killing effect of cytotoxic T lymphocyte（CTL） inducted by tumor-oaded UBDC.Results After induction of HUVEC and loading the tumor cell antigen,DC CD80,CD40,CD83,CD1a and MHC-II molecules expression were increased,generated CTL could specifically kill esophagal cancer cells EC9706,its killing rate to EC9706 had no statistical difference compared with that of CTL activated by cytokine-induced DC（P0.05）.Conclusion Using Transwell chamber primary HUVEC for inducing the differentiation of UBMC to DCs is a feasible experimental method.

关 键 词：树突状细胞 脐静脉内皮细胞 脐血单个核细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]