负载胶质瘤干细胞抗原的树突状细胞对脑胶质瘤治疗作用的研究

发布时间：2018-06-19 10:28

本文选题：胶质瘤干细胞 + 脑胶质瘤　；参考：《青岛大学》2014年硕士论文

【摘要】：目的体外培养大鼠C6细胞,从中分离、培养与鉴定胶质瘤干细胞,比较普通胶质瘤细胞及胶质瘤干细胞的致瘤性；由大鼠骨髓提取单核细胞诱导分化为树突细胞,分别用胶质瘤细胞、胶质瘤干细胞全细胞抗原致敏树突细胞制备疫苗；构建大鼠脑胶质瘤模型,经尾静脉分别注射两种树突细胞疫苗,观察大鼠生存情况并分析。方法体外常规培养C6细胞,取对数生长期细胞于无血清干细胞培养基中诱导培养脑胶质瘤干细胞,有限稀释法观察其增殖能力,并对其进行鉴定。分别将C6及胶质瘤干细胞接种于大鼠皮下,行PET-CT检查比较其致瘤性；提取大鼠骨髓源性单核细胞,加入细胞因子诱导成树突状细胞并对其鉴定,分别加入C6及胶质瘤干细胞冻融抗原,制备树突细胞疫苗；利用立体定向方法建立大鼠脑胶质瘤模型,观察其生存期、生活状态,肿瘤病理特点,行MRI检查观察肿瘤生长规律；分别将两种树突细胞疫苗由尾静脉注入脑内已成瘤大鼠体内,观察比较大鼠生存期,对生存情况进行分析。结果胶质瘤干细胞在无血清培养基中呈细胞球样悬浮生长,肿瘤干细胞标志物CD133, Nestin均阳性表达,具备自我更新及增殖能力,致瘤性强于普通胶质瘤细胞；树突细胞具有典型的树突状结构,特异性表面标志物OX62高表达；立体定向法构建大鼠脑胶质瘤模型成功率高,模型生存周期稳定,病理特点与人脑胶质瘤相似；注射利用胶质瘤干细胞抗原制备树突细胞疫苗大鼠生存时间长于普通树突细胞疫苗。结论胶质瘤干细胞致敏的树突细胞较普通胶质瘤细胞致敏的树突细胞有更强的抗肿瘤效果。
[Abstract]:Objective to isolate, culture and identify glioma stem cells from rat C6 cells in vitro, compare the tumorigenicity of normal glioma cells and glioma stem cells, and differentiate into dendritic cells from rat bone marrow derived monocytes. Glioma cells and glioma stem cells were used to sensitize dendritic cells, and rat glioma models were established. Two kinds of dendritic cell vaccines were injected via tail vein to observe the survival and analyze the survival of rats. Methods C6 cells were cultured in vitro and cultured in serum-free stem cell culture medium. The proliferative ability of C6 cells was observed by limited dilution method and identified. C6 and glioma stem cells were inoculated subcutaneously in rats. The tumorigenicity of C6 and glioma stem cells was compared by PET-CT. The dendritic cell vaccine was prepared by adding C6 and glioma stem cell freeze-thawed antigen respectively, the rat glioma model was established by stereotactic method, the survival time, living state, pathological characteristics of the tumor were observed, and the tumor growth rule was observed by MRI. Two kinds of dendritic cell vaccines were injected into the brain of tumor-forming rats from tail vein to observe the survival time of the rats and analyze the survival conditions. Results Glioma stem cells were cultured in serum-free medium, and the tumor stem cell markers CD133 and nestin were all positive. The glioma stem cells had the ability of self-renewal and proliferation, and the tumorigenicity of glioma cells was stronger than that of normal glioma cells. Dendritic cells have typical dendritic structure and high expression of specific surface marker OX62. Stereotactic method has a high success rate in the construction of rat glioma model, the survival cycle of the model is stable, and the pathological characteristics are similar to that of human glioma. The survival time of dendritic cell vaccine was longer than that of ordinary dendritic cell vaccine by injection of glioma stem cell antigen. Conclusion dendritic cells sensitized by glioma stem cells have stronger antitumor effect than those sensitized by normal glioma cells.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41

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