【摘要】：在胚胎發(fā)育過(guò)程中,脊索誘導(dǎo)其背側(cè)中線的外胚層(神經(jīng)外胚層)形成神經(jīng)板并發(fā)育成神經(jīng)管。緊接著神經(jīng)管閉合,管腔特化成腦室,背部區(qū)域特化成端腦。當(dāng)端腦起始發(fā)育的時(shí)候,組成神經(jīng)管的神經(jīng)上皮干細(xì)胞,不斷的增殖擴(kuò)增,逐漸轉(zhuǎn)變?yōu)榉派錉钅z質(zhì)前體細(xì)胞,最后這些膠質(zhì)前體細(xì)胞進(jìn)行遷移分化最終形成了大腦皮層。神經(jīng)管發(fā)育異常將導(dǎo)致一系列疾病。因此,利用多能性干細(xì)胞,發(fā)展一種靈長(zhǎng)類神經(jīng)管發(fā)育模型,能夠幫助我們研究神經(jīng)管疾病。本研究建立了一種穩(wěn)健的體系,該體系能夠在單細(xì)胞水平上進(jìn)行克隆擴(kuò)增并形成微小的類神經(jīng)管結(jié)構(gòu)：另外,單細(xì)胞不僅能夠在體外產(chǎn)生功能神經(jīng)元,還能在體內(nèi)存活并廣泛再生神經(jīng)元軸突：除此之外,本研究還發(fā)現(xiàn)神經(jīng)上皮干細(xì)胞特性的維持與類神經(jīng)管結(jié)構(gòu)的形成依賴高能量代謝：通過(guò)Notch信號(hào)與細(xì)胞粘附分子來(lái)調(diào)控Wnt信號(hào)通路,單個(gè)神經(jīng)上皮干細(xì)胞能夠轉(zhuǎn)變?yōu)榉派錉钅z質(zhì)前體細(xì)胞：最后,利用"NESC-TO-NTs"體系,本研究成功模擬了葉酸在神經(jīng)管閉合過(guò)程中的作用,表明葉酸可以調(diào)控多種機(jī)制預(yù)防神經(jīng)管疾病�？梢�(jiàn),本研究建立了一個(gè)能夠在體外研究神經(jīng)管發(fā)育和疾病理想的體系。大腦皮層是大腦中最復(fù)雜的結(jié)構(gòu)之一,其損傷會(huì)導(dǎo)致一系列的神經(jīng)發(fā)育和神經(jīng)退行性疾病。在結(jié)構(gòu)上,大腦皮層可以分為6層,第1層為cajal細(xì)胞層,2、3、4層為上層投射神經(jīng)元,5、6層為底層投射神經(jīng)元。根據(jù)神經(jīng)元的位置和突觸連接方式,5、6層又稱為離皮質(zhì)投射神經(jīng)元。離皮質(zhì)投射神經(jīng)元的分化方法缺乏,限制了研究其在大腦皮層中的發(fā)育和功能。本研究開(kāi)發(fā)了“兩步法”能夠從人胚胎干細(xì)胞快速誘導(dǎo)離皮質(zhì)神經(jīng)元：(1)誘導(dǎo)胚胎干細(xì)胞轉(zhuǎn)變?yōu)樯窠?jīng)上皮干細(xì)胞；(2)神經(jīng)上皮干細(xì)胞分化產(chǎn)生8096的高純度離皮質(zhì)投射神經(jīng)元。本方法獲取的神經(jīng)上皮干細(xì)胞不但能夠在單細(xì)胞水平進(jìn)行長(zhǎng)時(shí)間的自我更新并自我重組形成類神經(jīng)管結(jié)構(gòu),還能夠穩(wěn)定的分化為離皮質(zhì)投射神經(jīng)元和中間神經(jīng)元。另外,本研究還發(fā)現(xiàn)：移植的神經(jīng)上皮干細(xì)胞能成功整合到小鼠腦中,并分化為投射神經(jīng)元,從而建立有效的突觸連接和特殊的投射模式。因此,有效的產(chǎn)生離皮質(zhì)投射神經(jīng)元,能夠促進(jìn)人們了解大腦皮層的發(fā)育機(jī)制,并為細(xì)胞治療提供充足的儲(chǔ)備資源。
[Abstract]:During embryonic development, the notochord induces the ectoderm (neuroectoderm) of the dorsal midline to form a nerve plate and develop into a neural tube. Then the nerve tube is closed, the lumen becomes the ventricle, and the back area becomes the terminal brain. When the terminal brain begins to develop, the neural epithelial stem cells that make up the neural tube proliferate and expand, and gradually transform into radial glial precursor cells. Finally, these glial precursor cells migrate and differentiate and finally form the cerebral cortex. Dysplasia of the neural tube can lead to a series of diseases. Therefore, the use of pluripotent stem cells to develop a primate neural tube development model can help us to study neural tube diseases. This study established a robust system of cloning and amplification at the single cell level and the formation of tiny neuron-like structures: in addition, single cells not only produced functional neurons in vitro, Can also survive and regenerate a wide range of neuronal axons in the body: It was also found that the maintenance of neural epithelial stem cell properties and the formation of neuron-like tubular structures depended on high energy metabolism: Wnt signaling pathways were regulated by Notch signals and cell adhesion molecules. Single neural epithelial stem cells can be transformed into radial glial progenitor cells. Finally, using the "NESC-TO-NTs" system, we successfully simulated the role of folic acid in the process of neural tube closure. It is suggested that folic acid can regulate multiple mechanisms for the prevention of neural tube diseases. Thus, an ideal system for studying neurotubules and diseases in vitro has been established. The cerebral cortex is one of the most complex structures in the brain. Its damage can lead to a series of neurodevelopmental and neurodegenerative diseases. In structure, the cerebral cortex can be divided into six layers, the first layer is the cajal cell layer, the second layer is the upper layer, the third layer is the upper layer, and the 5 layer is the bottom layer. According to the location and synaptic connection of neurons, the 5th layer is also called the apocortical projective neurons. The lack of differentiation of apocortical projective neurons limits the development and function of these neurons in the cerebral cortex. In this study, a "two-step" method was developed to rapidly induce dissociated neurons from human embryonic stem cells: (1) inducing embryonic stem cells into neural epithelial stem cells; (2) the differentiation of neural epithelial stem cells (NSCs) produced 8096 high purity detached cortical projective neurons. The neural epithelial stem cells obtained by this method can not only self-renew and self-recombine for a long time at the single cell level to form neuron-like structures, but also can steadily differentiate into apocortical projective neurons and intermediate neurons. In addition, we also found that the transplanted neural epithelial stem cells could be successfully integrated into the mouse brain and differentiated into projective neurons, thus establishing effective synaptic connections and special projection patterns. Therefore, the effective production of apocortical projective neurons can promote the understanding of the development mechanism of cerebral cortex and provide sufficient resources for cell therapy.
【學(xué)位授予單位】：中國(guó)科學(xué)技術(shù)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q42

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