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猕猴胚胎干细胞的神经谱系分化及胶质祖细胞迁移机理研究
其他题名Neural Lineage Development of Rhesus Monkey Embryonic Stem Cells and the Mechanism of the Migration of Glial Precursors
陈洪伟
学位类型博士
导师季维智
2008
学位授予单位中国科学院研究生院
学位授予地点北京
关键词猕猴胚胎干细胞 神经谱系发育 No 迁移 Netrin-dcc信号
摘要由于伦理和材料来源的限制,目前对灵长类早期神经发育缺乏深入地了解。与啮齿类动物相比,猕猴在遗传和生理上与人类更接近,因此猕猴胚胎干细胞(rESCs)研究具有重要的研究价值,不仅能为研究发育生物学基础理论提供良好的模型,而且可为细胞替代性治疗提供大量的供体细胞。本文以rESCs为主要研究对象,在rESCs定向分化为神经细胞的基础上着重研究神经谱系分化及调控胶质祖细胞迁移的机理。主要结论如下:1) rESCs来源的神经上皮干/前体细胞(NEPs)主要变为辐射状胶质细胞(RG)后再通过中间类型的祖细胞——神经元祖细胞(NPs)和胶质祖细胞(GPs)——分别分化为神经元和胶质细胞。同时,NEPs/RG细胞群具有早期神经管背-腹和前-后轴空间特性。NEPs/RG的维持受Notch和FGFR信号作用。此外,实验中还纯化和鉴定了猕猴胶质限定性前体细胞(GRPs)。结果表明,rESCs的神经谱系分化能够模拟体内发育过程,并与啮齿类动物早期神经谱系变化过程相似。2) 气体信号分子NO(由10μM—250μM SNP供体释放)促进rESCs来源的A2B5+/Nestin/PSA-NCAM胶质祖细胞迁移。进一步研究发现Netrin-DCC信号通路介导了NO启动的细胞迁移过程。同时,Ca2也参与调控胶质祖细胞的迁移。此外,细胞外基质和整合素α6亦可能与Netrin-DCC相互作用调控细胞迁移。结果显示,NO通过激活一个复杂的信号网络系统调控胶质祖细胞迁移。本实验的研究结果有助于揭示灵长类中枢神经系统发育的机理,同时也能为治疗神经系统退行性疾病提供阶段特异性的供体细胞。
其他摘要So far the knowledge on the early neural development in primates is not well understood due to the restrictions of ethics and material sources. Rhesus monkey embryonic stem cells (rESCs) are ideal candidates for basic research and generation of donor cells for transplantation therapies because the monkey takes advantages over rodents in close genetic and physiological relation to human. This study mainly focused on the neural lineage development with rESCs and the mechanism regulating the migration of glial precursors (GPs). The results are shown as follows: 1. The neuroepithelial stem/progenitor cells (NEPs) derived from rESCs predominantly switched into radial glia (RG) and differentiated into neurons and glia via intermediate neuronal precursors (NPs) and GPs, respectively. Moreover, NEPs/RG pool displayed spatial identity of both dorsoventral and anteroposterior axes of the early neural tube. The maintenance of NEPs/RG was regulated by Notch and FGFR signaling. Additionally, glial restricted precursors (GRPs) were purified, and characterized in vitro and in vivo. The results suggested that the early neural lineage differentiation with rESCs in vitro could essentially mimic the in vivo process and was quite similar to the lineage development in rodents. 2. A2B5+/Nestin-/PSA-NCAM- GPs derived from rESCs initiated to migrate in vitro after NO (released from 10μM—250μM SNP) addition with time-lapse video imaging. Further investigation revealed that Netrin-DCC signaling with Ca2+ involved mediated the cell migration. Besides, Netrin-DCC signaling might combine with extracellualr matrix (ECM) and integrin α6 to cooperatively regulate the migration. It was suggested that NO regulated the migration of the GPs by activating a complex network including Netrin-DCC, Ca2+, ECM and integrin signalings. This study would facilitate to understand the molecular mechanism of the central nervous system (CNS) development in primates and to provide stage-specific donor cells for therapy on degenerative diseases in the CNS.
语种中文
文献类型学位论文
条目标识符http://ir.kiz.ac.cn/handle/152453/6455
专题生殖与发育生物学
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GB/T 7714
陈洪伟. 猕猴胚胎干细胞的神经谱系分化及胶质祖细胞迁移机理研究[D]. 北京. 中国科学院研究生院,2008.
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