新生大鼠神經(jīng)干細胞定向分化為少突膠質(zhì)細胞的實驗研究
發(fā)布時間：2018-09-13 13:14
【摘要】： 第一部分新生大鼠神經(jīng)干細胞的分離、培養(yǎng) 研究背景及目的神經(jīng)干細胞(neural stem cells, NSCs)是多分化潛能的細胞,可分化為神經(jīng)系統(tǒng)的神經(jīng)元、星形膠質(zhì)細胞和少突膠質(zhì)細胞,在胚胎及成年哺乳類動物中廣泛存在。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,采用體外無血清懸浮培養(yǎng),以獲得能在體外長期生存的NSC;通過單細胞克隆實驗檢測其自我更新能力,免疫細胞化學(xué)檢測NSC標志物nestin表達及分化為神經(jīng)元和膠質(zhì)細胞的能力;用細胞計數(shù)法研究其生長特點并繪制生長曲線。 結(jié)果獲得的NSC能在體外長期生存,具有很強的增殖能力、自我更新能力,能分化為神經(jīng)元和膠質(zhì)細胞,具有多向分化能力,表達nestin,體外傳代至10代細胞數(shù)量擴增了40倍。 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期
[Abstract]:The first part: isolation and culture of neural stem cells from neonatal rats background and objective Neural stem cells (neural stem cells, NSCs) are multi-differentiated cells. The isolation and culture of neurons, astrocytes and oligodendrocytes, which can differentiate into nervous system, are the premise of the basic and applied research on the separation and culture of .NSC in embryos and adult mammals. The aim of this part is to obtain NSCs, from neonatal rats and lay a foundation for further study of its proliferation and differentiation. Methods NSC, was isolated from rat telencephalon and midbrain within 24 hours and cultured in vitro with serum-free suspension. NSC;, which could survive for a long time in vitro, was tested for its self-renewal ability by single cell clone assay. Immunocytochemistry was used to detect the expression of NSC marker nestin and its ability to differentiate into neurons and glial cells. Results the obtained NSC could survive for a long time in vitro, had strong ability of proliferation, self-renewal, differentiation into neurons and glial cells, and had the ability of multi-directional differentiation. The number of cells subcultured to the 10th passage of the expressed nestin, in vitro was expanded by 40 times. Conclusion\
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R329.28
[Abstract]:The first part: isolation and culture of neural stem cells from neonatal rats background and objective Neural stem cells (neural stem cells, NSCs) are multi-differentiated cells. The isolation and culture of neurons, astrocytes and oligodendrocytes, which can differentiate into nervous system, are the premise of the basic and applied research on the separation and culture of .NSC in embryos and adult mammals. The aim of this part is to obtain NSCs, from neonatal rats and lay a foundation for further study of its proliferation and differentiation. Methods NSC, was isolated from rat telencephalon and midbrain within 24 hours and cultured in vitro with serum-free suspension. NSC;, which could survive for a long time in vitro, was tested for its self-renewal ability by single cell clone assay. Immunocytochemistry was used to detect the expression of NSC marker nestin and its ability to differentiate into neurons and glial cells. Results the obtained NSC could survive for a long time in vitro, had strong ability of proliferation, self-renewal, differentiation into neurons and glial cells, and had the ability of multi-directional differentiation. The number of cells subcultured to the 10th passage of the expressed nestin, in vitro was expanded by 40 times. Conclusion\
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R329.28
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