Aszl在小鼠胚胎干细胞向原始生殖细胞特化中的功能研究

发布时间：2018-01-28 19:44

本文关键词： 胚胎干细胞原始生殖细胞 Asz1　出处：《华东师范大学》2012年硕士论文　论文类型：学位论文

【摘要】：胚胎干细胞具有自我更新的能力,并且可以分化形成身体的各种组织细胞。胚胎干细胞研究对于阐明人类以及哺乳动物的生命规律具有重大的意义,已经成为当今生命科学和医学研究最为关注的热点领域。作为延续生命的要素之一,生殖细胞在胚胎发育过程中起着重要的作用。胚胎干细胞在体外通过悬滴培养和后续摇动培养的方式可形成具有三胚层结构的拟胚体,一定程度上在体外模拟体内胚胎发育的过程。我们以胚胎干细胞为技术平台,建立了研究生殖细胞分化所需的多种细胞模型,研究胚胎干细胞向生殖细胞诱导分化的转录调控网络。我们根据文献筛选了一批在生殖细胞有特异表达的基因,通过病毒包装感染的方式,建立了一批特定基因过量表达和低表达的稳定胚胎干细胞系,在这些特定基因过量表达和低表达的胚胎干细胞系中通过拟胚体的诱导分化模型,在不同时间点检测SSEA1+和碱性磷酸酶高活性的原始生殖细胞的形成效率,发现了Aszl过量表达具有促进SSEA1阳性群体比例升高和碱性磷酸酶活性染色阳性克隆增多的功能,而Asz1低表达又具有能够使碱性磷酸酶活性染色阳性克隆减少的功能。另外,我们的实时定量RT-PCR结果显示Asz1过量表达后,一些生殖细胞特异表达的基因如Daz1, Vasa, Oct4等的表达量都比对照组升高,而Asz1低表达后,这些生殖细胞特异表达的基因如Daz1, Vasa, Oct4等的表达量都比对照组降低。综上所述,Asz1可能在鼠胚胎干细胞中具有促进原始生殖细胞生成的功能。
[Abstract]:Embryonic stem cells have the ability to self-renew and can differentiate into various tissue cells of the body. Embryonic stem cell research is of great significance to clarify the laws of human and mammalian life. It has become a hot area of life science and medical research, as one of the elements of life. Germ cells play an important role in the process of embryonic development. Embryonic stem cells can form embryoid bodies with three-layer structure through suspension culture and subsequent shaking culture in vitro. To some extent, the process of embryonic development in vitro is simulated. We have established a variety of cell models for the study of germ cell differentiation based on embryonic stem cells. To study the transcriptional regulatory network of embryonic stem cell differentiation into germ cells, we have screened a number of genes specifically expressed in germ cells according to the literature, and infected by virus packaging. A number of stable embryonic stem cell lines with overexpression and low expression of specific genes were established and induced by embryoid differentiation in these specific gene overexpression and low expression embryonic stem cell lines. The formation efficiency of primordial germ cells with high activity of SSEA1 and alkaline phosphatase was detected at different time points. It was found that overexpression of Aszl could increase the proportion of SSEA1 positive population and the number of ALP positive clones. However, the low expression of Asz1 can reduce the expression of ALP positive clones. In addition, our real-time quantitative RT-PCR results showed that Asz1 was overexpressed. Some genes specifically expressed in germ cells, such as Daz1, Vasa, Oct4 and so on, were higher than those of the control group, but the expression of Asz1 was lower than that of the control group. The expression of these germ cell specifically expressed genes such as Daz1, Vasa, Oct4 were lower than that of the control group. Asz1 may have the function of promoting primordial germ cell formation in mouse embryonic stem cells.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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