本文選題：神經(jīng)干細(xì)胞　切入點(diǎn)：新生大鼠　出處：《重慶醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 第一部分新生大鼠神經(jīng)干細(xì)胞的分離、培養(yǎng) 研究背景與目的神經(jīng)干細(xì)胞(neural stem cell, NSC)是近年來(lái)神經(jīng)科學(xué)領(lǐng)域的研究熱點(diǎn)。NSC是具有自我更新能力和多向分化潛能的細(xì)胞,可分化為神經(jīng)系統(tǒng)的神經(jīng)元、星形膠質(zhì)細(xì)胞和少突膠質(zhì)細(xì)胞,在胚胎及成年哺乳類動(dòng)物中廣泛存在。NSC的分離培養(yǎng)是其基礎(chǔ)和應(yīng)用研究的前提,本部分目的在于獲得新生大鼠NSCs,為深入研究其增殖分化奠定基礎(chǔ)。 方法從新生24h內(nèi)大鼠端腦及中腦部位分離NSC,采用體外無(wú)血清懸浮培養(yǎng),以獲得能在體外長(zhǎng)期生存的NSC;通過(guò)免疫細(xì)胞化學(xué)檢測(cè)NSC標(biāo)記物nestin表達(dá)及分化為神經(jīng)元和膠質(zhì)細(xì)胞的能力。 結(jié)果獲得的NSC能在體外長(zhǎng)期生存,具有很強(qiáng)的增殖能力、自我更新能力,表達(dá)nestin,具有多向分化能力,能分化為神經(jīng)元和膠質(zhì)細(xì)胞。 結(jié)論成功地從新生大鼠端腦和中腦中分離得到了能在體外長(zhǎng)期培養(yǎng)的NSC,培養(yǎng)的NSC表達(dá)nestin,有自我更新、多向分化的特點(diǎn),通過(guò)培養(yǎng)細(xì)胞大量擴(kuò)增。 第二部分ATRA、NGF誘導(dǎo)新生大鼠神經(jīng)干細(xì)胞定向分化為膽堿能神經(jīng)元的實(shí)驗(yàn)研究 研究背景及目的神經(jīng)干細(xì)胞為神經(jīng)系統(tǒng)疾病細(xì)胞替代治療提供了細(xì)胞來(lái)源,疾病的治療需要大量特定表型和功能的神經(jīng)細(xì)胞,因此,對(duì)NSC分化為某種特定表型的研究成為神經(jīng)科學(xué)研究的焦點(diǎn)。阿爾茨海默病(AD)是一種以基底前腦膽堿能神經(jīng)元的退變?yōu)橹饕±硖卣鞯纳窠?jīng)系統(tǒng)變性疾病,補(bǔ)充膽堿能神經(jīng)元是治療該疾病的一種策略。本部分的目的在于在體外環(huán)境下使NSC定向分化為膽堿能神經(jīng)元,為深入研究NSC體外分化條件,探討誘導(dǎo)分化機(jī)制,提高膽堿能神經(jīng)元分化效能奠定細(xì)胞學(xué)基礎(chǔ)。 方法在第3代NSCs無(wú)血清培養(yǎng)體系中加入ATRA和/或NGF,分別對(duì)NSC克隆進(jìn)行ATRA、NGF、ATRA聯(lián)合NGF誘導(dǎo)分化,通過(guò)免疫細(xì)胞學(xué)方法檢測(cè)膽堿能神經(jīng)元標(biāo)志物CHAT的表達(dá),比較不同誘導(dǎo)因素對(duì)NSC定向分化為膽堿能神經(jīng)元的作用。 結(jié)果ATRA、NGF單獨(dú)誘導(dǎo)均提高CHAT陽(yáng)性細(xì)胞表達(dá),與血清對(duì)照組比較有顯著性差異(P0.05),而ATRA聯(lián)合NGF誘導(dǎo)組與單獨(dú)應(yīng)用NGF組比較不能顯著提高CHAT陽(yáng)性細(xì)胞率,二者無(wú)顯著性差異(P0.05)。 結(jié)論ATRA和NGF均有誘導(dǎo)NSC向膽堿能神經(jīng)元分化的作用,但兩者并不存在協(xié)同誘導(dǎo)作用。
[Abstract]:Part 1 Isolation and culture of neural stem cells from neonatal rats
Background and objective neural stem cells (neural stem cell, NSC) is a hotspot in the field of neuroscience is.NSC with self-renewal and multilineage differentiation potential of cells can differentiate into neurons, astrocytes and oligodendrocytes in embryonic and adult mammalian animal widely existed in culture.NSC separation is the premise of basic and applied research. The objective of this part is to obtain the NSCs neonatal rat, which laid the foundation for the further study on its proliferation and differentiation.
Methods NSC was isolated from the midbrain and midbrain of neonatal 24h in rats, and serum-free suspension culture was used to obtain long-term NSC in vitro. The expression of NSC marker nestin and the ability to differentiate into neurons and glial cells were detected by immunocytochemistry.
Results NSC can survive in vitro for a long time. It has strong proliferative ability, self-renewal ability, and nestin expression. It has multiple differentiation ability and can differentiate into neurons and glial cells.
Conclusion NSC, which can be cultured in vitro for a long time, is successfully isolated from the midbrain and midbrain of neonatal rats. The NSC expression of nestin is self renewing and multidirectional. It is amplified by culture cells.
The second part ATRA and NGF induced the directional differentiation of neural stem cells into cholinergic neurons in neonatal rats
Background and objective neural stem cells provide a cell source for cell replacement therapy of nervous system diseases, disease treatment requires a lot of specific phenotype and function of nerve cells, therefore, to study the differentiation of NSC into a specific phenotype has become the focus of neuroscience research. Alzheimer's disease (AD) is a neurodegenerative disease in basal forebrain cholinergic neurons degeneration is the main pathological features of cholinergic neurons, is a kind of strategy in the treatment of the disease. The purpose of this section is in vitro under the environment of NSC to differentiate into cholinergic neurons, for further research on NSC in vitro differentiation conditions, explore the mechanism of inducing differentiation, improve cholinergic cell basis for the neuronal differentiation efficiency.
Methods with ATRA and / or NGF system in serum culture in the third generation NSCs, respectively NSC clones were ATRA, NGF, ATRA and NGF to induce differentiation, by immunocytologic detection method of cholinergic expression of neuronal markers CHAT, comparison of different factors on NSC induced differentiation into cholinergic neurons.
Results ATRA and NGF alone increased the expression of CHAT positive cells. There was a significant difference compared with the serum control group (P0.05), while the ATRA plus NGF induction group did not significantly increase the CHAT positive cells rate compared with the NGF group alone, but there was no significant difference between the two groups (P0.05).
Conclusion both ATRA and NGF can induce the differentiation of NSC to cholinergic neurons, but there is no synergistic effect between them.

【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R329.28

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 杜海燕;NeuroD對(duì)短暫缺氧性腦損傷模型小鼠的學(xué)習(xí)記憶功能及其腦內(nèi)神經(jīng)干細(xì)胞分化作用的影響[D];福建醫(yī)科大學(xué);2013年

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本文編號(hào)：1717407