本文選題：大鼠　切入點：神經(jīng)干細胞　出處：《鄭州大學(xué)》2006年碩士論文

【摘要】：目的：建立新生大鼠嗅球神經(jīng)干細胞(neural stem cells，NSCs)體外分離培養(yǎng)方法。通過對不同發(fā)育階段大鼠嗅球NSCs體外培養(yǎng)的形態(tài)學(xué)觀察及免疫熒光化學(xué)染色結(jié)果分析，探討不同發(fā)育階段大鼠嗅球NSCs增殖、分化特性。使用不同濃度的胎牛血清、IGF-I和BDNF誘導(dǎo)大鼠嗅球NSCs的分化，探討促進大鼠嗅球NSCs向神經(jīng)元分化的因素，為大鼠嗅球NSCs的深入研究與臨床應(yīng)用打基礎(chǔ)。 材料與方法：從新生大鼠嗅球組織中分離NSCs，采用神經(jīng)細胞球形成法，以無血清培養(yǎng)液，使用生長因子(bFGF、EGF)，進行體外擴增培養(yǎng)，以機械分離法連續(xù)傳代，相差顯微鏡及電子顯微鏡觀察細胞形態(tài)，免疫細胞化學(xué)法檢測Nestin表達，四甲基偶氮唑鹽(MTT)比色法和流式細胞儀檢測大鼠嗅球NSCs的增殖能力。取胎齡18天(embryonic day 18，ED18)、出生24h以內(nèi)(postnatal within 24h，P0)及2月齡(2 month，M2)的大鼠嗅球組織行單細胞克隆，免疫熒光化學(xué)染色鑒定嗅球NSCs，，并計數(shù)分化為神經(jīng)元、星形膠質(zhì)細胞和少突膠質(zhì)細胞的比例。用免疫細胞化學(xué)染色和流式細胞儀分選細胞法觀察不同濃度的胎牛血清、IGF-I和(或)BDNF對嗅球NSCs分化為神經(jīng)元、星型膠質(zhì)細胞、少突膠質(zhì)細胞的影響。 結(jié)果：光鏡下原代培養(yǎng)見細胞呈大小不一、懸浮生長的細胞團，邊界清晰，折光性好，細胞團中細胞排列較為致密，傳代培養(yǎng)的細胞形態(tài)特點與原代培
[Abstract]:Objective: to establish a method of isolation and culture of neural stem cells from olfactory bulb neural stem cells (NSCs) of newborn rats in vitro. The morphological observation and immunofluorescence staining results of NSCs in olfactory bulb of rats at different stages of development were observed. To investigate the proliferation and differentiation characteristics of olfactory bulb NSCs in rats at different developmental stages, the differentiation of olfactory bulb NSCs was induced by different concentrations of fetal bovine serum IGF-I and BDNF, and the factors that promoted the differentiation of rat olfactory bulb NSCs into neurons were discussed. To lay a foundation for the further study and clinical application of rat olfactory bulb NSCs. Materials and methods: NSCs were isolated from the olfactory bulb of newborn rats. NSCs were isolated from the olfactory bulb tissue of newborn rats. The cells were cultured in vitro using serum-free medium and growth factor BFGFG EGFN. The cells were subcultured by mechanical separation method. Phase contrast microscope and electron microscope were used to observe cell morphology and immunocytochemistry method was used to detect the expression of Nestin. The proliferative ability of rat olfactory bulb NSCs was detected by MTT colorimetry and flow cytometry. Single cell clones were performed in the olfactory bulb tissues of rats with embryonic day 18 (ED18), postnatal within 24 h (P0) and 2-month old (2 monththophane M2) at gestational age of 18 days. NSCs of olfactory bulb were identified by immunofluorescence staining and differentiated into neurons. The ratio of astrocytes to oligodendrocytes. Immunocytochemical staining and flow cytometry were used to observe the differentiation of NSCs from olfactory bulb to neurons and astrocytes in fetal bovine serum of different concentrations. The effect of oligodendrocytes. Results: under the light microscope, the cells in primary culture were different in size and size. The cell clusters with suspending growth had clear boundary, good refraction and compact arrangement of cells in the cell clusters. The morphological characteristics and primary culture of the cells in passage culture were similar to those in the primary culture.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R329

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