本文選題：膠質(zhì)瘤干細(xì)胞 + 腦膠質(zhì)瘤��； 參考：《青島大學(xué)》2014年碩士論文

【摘要】：目的體外培養(yǎng)大鼠C6細(xì)胞,從中分離、培養(yǎng)與鑒定膠質(zhì)瘤干細(xì)胞,比較普通膠質(zhì)瘤細(xì)胞及膠質(zhì)瘤干細(xì)胞的致瘤性；由大鼠骨髓提取單核細(xì)胞誘導(dǎo)分化為樹突細(xì)胞,分別用膠質(zhì)瘤細(xì)胞、膠質(zhì)瘤干細(xì)胞全細(xì)胞抗原致敏樹突細(xì)胞制備疫苗；構(gòu)建大鼠腦膠質(zhì)瘤模型,經(jīng)尾靜脈分別注射兩種樹突細(xì)胞疫苗,觀察大鼠生存情況并分析。 方法體外常規(guī)培養(yǎng)C6細(xì)胞,取對數(shù)生長期細(xì)胞于無血清干細(xì)胞培養(yǎng)基中誘導(dǎo)培養(yǎng)腦膠質(zhì)瘤干細(xì)胞,有限稀釋法觀察其增殖能力,并對其進(jìn)行鑒定。分別將C6及膠質(zhì)瘤干細(xì)胞接種于大鼠皮下,行PET-CT檢查比較其致瘤性；提取大鼠骨髓源性單核細(xì)胞,加入細(xì)胞因子誘導(dǎo)成樹突狀細(xì)胞并對其鑒定,分別加入C6及膠質(zhì)瘤干細(xì)胞凍融抗原,制備樹突細(xì)胞疫苗；利用立體定向方法建立大鼠腦膠質(zhì)瘤模型,觀察其生存期、生活狀態(tài),腫瘤病理特點(diǎn),行MRI檢查觀察腫瘤生長規(guī)律；分別將兩種樹突細(xì)胞疫苗由尾靜脈注入腦內(nèi)已成瘤大鼠體內(nèi),觀察比較大鼠生存期,對生存情況進(jìn)行分析。 結(jié)果膠質(zhì)瘤干細(xì)胞在無血清培養(yǎng)基中呈細(xì)胞球樣懸浮生長,腫瘤干細(xì)胞標(biāo)志物CD133, Nestin均陽性表達(dá),具備自我更新及增殖能力,致瘤性強(qiáng)于普通膠質(zhì)瘤細(xì)胞；樹突細(xì)胞具有典型的樹突狀結(jié)構(gòu),特異性表面標(biāo)志物OX62高表達(dá)；立體定向法構(gòu)建大鼠腦膠質(zhì)瘤模型成功率高,模型生存周期穩(wěn)定,病理特點(diǎn)與人腦膠質(zhì)瘤相似；注射利用膠質(zhì)瘤干細(xì)胞抗原制備樹突細(xì)胞疫苗大鼠生存時(shí)間長于普通樹突細(xì)胞疫苗。 結(jié)論膠質(zhì)瘤干細(xì)胞致敏的樹突細(xì)胞較普通膠質(zhì)瘤細(xì)胞致敏的樹突細(xì)胞有更強(qiáng)的抗腫瘤效果。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R739.41

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本文編號：2039579