冻存后脐血DC介导的食管癌疫苗体内致瘤性研究  

Oncogenicity of the vaccine of esophageal cancer cells fused with dendritic cells derived from the umbilical cord blood after cryopreservation in vivo

作　　者：郭光华[1] 谢丽华[1] 陈素钻[1] 俞晶[1] 张娟[1] 罗丽莉[1] 苏中静[1] 董红梅[1] 徐虹[1] 吴利标[1] 

机构地区：[1]汕头大学医学院第一附属医院消化内科,515041

出　　处：《实用医学杂志》2006年第4期367-369,共3页The Journal of Practical Medicine

基　　金：广东省自然科学基金资助项目(编号:021228)

摘　　要：目的:探讨低温冻存后脐血树突状细胞(DC)与EC109食管癌细胞融合疫苗体内致瘤性。方法:免疫磁珠标记法分选人脐血CD34+干细胞,细胞因子扩增培养为成熟DC,聚二乙醇(PEG)融合法制备EC109-DC融合疫苗,流式细胞术双阳性标记鉴定融合疫苗,-80℃低温冰箱冻存,3周后融冻,流式细胞术再次鉴定融合疫苗,行体内致瘤性研究。结果:(1)融冻后疫苗体外半悬浮生长;同时高表达FR和CD80;(2)融冻后疫苗接种于SCID小鼠体内未见肿瘤形成,脾、肺和肝等器官未出现肿瘤病变。结论:冻存未破坏融合疫苗的完整性,融冻瘤苗无体内致瘤性,其亲本EC109细胞的恶性生长特征已丧失,安全可靠。Objective To study the oncogenicity of the vaccine of esophageal cancer cells fused with dendritic cells （DC） derived from the umbilical cord blood after cryopreservation in vivo. Methods CD34^＋ hematopoietic stem cells were isolated from umbilical cord blood using CD34^＋ Progenitor Cell Isolation Kit by magnetic cell sorting system （MACS）, and then the CD34^＋ cells were expanded with cytokines as DCs, and fused with EC109 cells by PEG3600, the fused cells were frozen and stored at -80℃ for three weeks. Their immunophenotypes of both the flash vaccine and the cryopreservated vaccine were assayed by flow cytometry. Oncogenicity of the vaccine of esophageal cancer cells fused with DC derived from the umbilical cord blood after cryopreservation were tested in vivo. Results （1） The fusion cells showed half-suspension growth and highly expressed not only FR but also CD80, whereas EC109 highly expressed only FR（87.37% ） and DCs highly expressed only CD80（82.36% ）. （2）No tumor was found in spleen, lung and liver after the fusion vaccine injection. Conclusions Cryopreservation does not cause significant changes in the phenotypes of the fused vaccine cells. The fusion vaccine after thawing have no oneogenicity in vivo.

关 键 词：癌症疫苗 树突细胞 低温保存 致瘤性试验 

分 类 号：R735.1[医药卫生—肿瘤]