胶质瘤干细胞的分离鉴定与生物学特性研究-微生物学专业论文.docxVIP

．支 ．支 lIII lIII I I III IIII I IIII I III 、t 1 9431 98 摘要 摘要 胶质瘤(Glioma)是起源于神经外胚层的肿瘤，发病率在颅内肿瘤中最高， 是严重危害人们生命健康的恶性肿瘤之一。但由于胶质瘤的浸润性、无限增生 等特点，目前临床上很难做到完全治愈，复发率较高。人们认为胶质瘤干细胞 (Glioma Stem cell，GSC)的存在，是胶质瘤难以根治的根源。胶质瘤干细胞应 该就是胶质瘤的“种子”细胞，这类细胞对胶质瘤的发生、转移和复发起着非常关 键的作用。 本研究利用无血清的培养基分离并培养了胶质瘤干细胞，以该类细胞为主 要研究对象，通过细胞免疫染色和流式细胞术对其进行了细胞生物学特性研究。 此外，还通过Real-time PCR进一步分析了胶质瘤干细胞的基因表达情况，探讨 了其分子生物学特性。 实验结果表明，本文通过无血清培养方法成功培养出了能形成克隆球，具 有自我更新能力和多向分化潜能的干细胞样细胞，且与正常神经干细胞不同， 该类细胞可以分化产生GFAP和Tujl共表达的异常细胞类型。此外，通过real-time PCR结果证实该类细胞比胶质瘤细胞具有更强的抗凋亡能力。通过本次研究，进 一步加深了对胶质瘤干细胞生物特性的认识，为胶质瘤发生、发展的机制的研 究提供了良好的细胞模型，希望有助于新的针对胶质瘤干细胞的治疗方法的研 发，为胶质瘤的早日治愈提供理论依据。 关键词：胶质瘤干细胞； 分离鉴定；生物特性分析 ABSTRACTABSTRACT ABSTRACT ABSTRACT Glioma,one of the most lethal cancers，originated from the neuroderm，has the highest incidence of intracranial tumors and is seriOUS harmful to people’S lives． However,due to the invasion and unlimited proliferation，glioma晰m the high recurrence rate is very difficult to cure，and the recurrence tumor is more malignant and more difficult to cure than primary．So people assulne that glioma stem cells (GSCs)，may be the true source of glioma difficult to cure．GSCs should be the‘‘seed’’ cells，playing a key role in tumor initiation，metastasis and relapse． In this study,we successfully isolated and cultured glioma stem cells with the sertllTl-free medium，and then we analyzed the characteristics of these cells by immunostaining and flow cytometry．Based on these，we also analyzed the gene expression of these cells through real—time PCR． Our results showed that we successfully isolated and cultured stem cell like cells in SFM medium，which Can forme tumor spheres and had the self-renewal and differentiation capacity．Through the cellular and molecular analysis，we inferred that these cells were the glioma stem cells．We hope this study will help to deepen understanding the characteristics of GSCs，provide a good model for the mechanism study of glioma and contribute to the development of