本文關(guān)鍵詞： 胚胎干細(xì)胞 原始生殖細(xì)胞 Asz1　出處：《華東師范大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：胚胎干細(xì)胞具有自我更新的能力,并且可以分化形成身體的各種組織細(xì)胞。胚胎干細(xì)胞研究對于闡明人類以及哺乳動物的生命規(guī)律具有重大的意義,已經(jīng)成為當(dāng)今生命科學(xué)和醫(yī)學(xué)研究最為關(guān)注的熱點(diǎn)領(lǐng)域。作為延續(xù)生命的要素之一,生殖細(xì)胞在胚胎發(fā)育過程中起著重要的作用。胚胎干細(xì)胞在體外通過懸滴培養(yǎng)和后續(xù)搖動培養(yǎng)的方式可形成具有三胚層結(jié)構(gòu)的擬胚體,一定程度上在體外模擬體內(nèi)胚胎發(fā)育的過程。我們以胚胎干細(xì)胞為技術(shù)平臺,建立了研究生殖細(xì)胞分化所需的多種細(xì)胞模型,研究胚胎干細(xì)胞向生殖細(xì)胞誘導(dǎo)分化的轉(zhuǎn)錄調(diào)控網(wǎng)絡(luò)。我們根據(jù)文獻(xiàn)篩選了一批在生殖細(xì)胞有特異表達(dá)的基因,通過病毒包裝感染的方式,建立了一批特定基因過量表達(dá)和低表達(dá)的穩(wěn)定胚胎干細(xì)胞系,在這些特定基因過量表達(dá)和低表達(dá)的胚胎干細(xì)胞系中通過擬胚體的誘導(dǎo)分化模型,在不同時(shí)間點(diǎn)檢測SSEA1+和堿性磷酸酶高活性的原始生殖細(xì)胞的形成效率,發(fā)現(xiàn)了Aszl過量表達(dá)具有促進(jìn)SSEA1陽性群體比例升高和堿性磷酸酶活性染色陽性克隆增多的功能,而Asz1低表達(dá)又具有能夠使堿性磷酸酶活性染色陽性克隆減少的功能。另外,我們的實(shí)時(shí)定量RT-PCR結(jié)果顯示Asz1過量表達(dá)后,一些生殖細(xì)胞特異表達(dá)的基因如Daz1, Vasa, Oct4等的表達(dá)量都比對照組升高,而Asz1低表達(dá)后,這些生殖細(xì)胞特異表達(dá)的基因如Daz1, Vasa, Oct4等的表達(dá)量都比對照組降低。綜上所述,Asz1可能在鼠胚胎干細(xì)胞中具有促進(jìn)原始生殖細(xì)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.
【學(xué)位授予單位】：華東師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329

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