中文摘要：

目的比较胶质瘤干细胞样抗原与非干细胞样抗原致敏树突状细胞（DCs）疫苗对胶质瘤的体外作用。方法体外用无血清法和血清法培养恶性胶质瘤细胞，有限稀释法检测其克隆形成率；冻融法获取相应胶质瘤抗原；GM—CSF、IL-4体外诱导人外周血单核细胞获取DCs，与抗原孵育后再激活T淋巴细胞，同位素法检测效应细胞对胶质瘤细胞的杀伤率；流式细胞术检测DCs表面分子变化以及非贴壁细胞中T细胞比例的变化。结果无血清培液中的胶质瘤细胞具有更高的克喹形成率（P〈0．01）；DCs经不同抗原刺激后HLA—A、HLA—DR、CD80、CD86表达上调（P〈0．01）；非贴壁细胞与DCs共培养后T细胞比例明显增高；干细胞样抗原激活的效应细胞对非干细胞阵细胞和干细胞样细胞杀伤率分别为（70，2±5，13）％和（56．7±7．81）％（P〈0．001），而非干细胞洋抗原活化的效应细胞相应的杀伤率为（36．6±6．45）％和（9．05±3．49）％（P〈0．001）。结论无血清培养液中的胶质瘤细胞具有更强的增殖能力，胶质瘤干细胞样抗原较传统抗原具有更强的抗原性，为进一步提高胶质瘤DCs疫苗疗效奠定基础。

英文摘要：

Objective To compare the different effects between DC -based vaccine implused by tumor stem - like cells （TSCs） and non - TSCs on cytotoxicity in vitro. Method Malignant glioma cell lines（ SHG62,SHG66, U87） were cultured in serum contained medium（SCM, DMEM with 10% FBS） or in serum free medium （SFM, DMEM/F12 supplemented with EGF/bFGF）respectively. Clone formation was observed by limiting dilution analysis. Tumor lysates were extracted from cells cultured in the two different mediums after three freeze - thaw cycles. Corresponsive DCs from PBMCs were co - cultured in RPMI1640 with GM - CSF and IL - 4, and then matured by tumor lysates to stimulate autologous T cells. HLA - A, HLA -DR,CD80, CD86 of DCs before and after maturation were checked by flowcytometry, and effector cells were checked with CD3, CD16 and CD56. Then effector cells and targeted cells were cocuhured with different ratio and cytotoxicity was investigated on scintillater after impregnation with thymidine labeled with tritium. Results Ceils cultured in the two mediums displayed different morphology. The cells in SEM had the ability of better self - renewal. DCs were successfully matured by tumor stem - like associated antigens. Flowcytometric analysis determined that HLA - A, HLA - DR, CD80, CD86 of DCs were up - regulated,and CD3 ＋ T cell ratio was significantly improved after co -cultured with corresponsive DCs （ P 〈 0. 01 ）. Effect cells loaded with tumor stem -like cells associated antigens killed （70. 2± 5. 13 ）% of non stem - like cells and （ 56. 7 ± 7.81 ） % of stem - like cells （ P 〈 0. 001 ） , however effector cells loaded with non stem - like ceils associated antigens eliminated only （ 36. 6± 6.45 ） % of non stem - like cells and （9. 05±3.49） % of stem- like ceils （ P 〈 0. 001 ）. Conclusions The glioma ceils in SFM had better proliferation, and DCs implused by tumor stem - like cells associated antigens augment the protection against malignant glioma cells in vitro, providing robus