本文選題：骨髓基質(zhì)干細(xì)胞　切入點：分離純化　出處：《蘇州大學(xué)》2007年碩士論文

【摘要】： 第一部分大鼠骨髓基質(zhì)干細(xì)胞的體外分離、培養(yǎng)及定向誘導(dǎo)分化 目的:探索大鼠骨髓基質(zhì)干細(xì)胞(bone marrow stromal cells, BMSCs)體外分離、培養(yǎng)及純化的合適實驗條件,并探討其在體外定向誘導(dǎo)分化為神經(jīng)元樣細(xì)胞的可行性。 方法:通過密度梯度離心法從成年大鼠骨髓中分離出BMSCs,而后通過貼壁培養(yǎng)法培養(yǎng)并進一步純化,觀察其生長特性;對純化后的BMSCs使用堿性成纖維生長因子(bFGF)和表皮生長因子(EGF)進行定向誘導(dǎo)分化,并進行免疫細(xì)胞化學(xué)染色鑒定分化結(jié)果。 結(jié)果:體外培養(yǎng)的BMSCs傳4代后,純度最高,可達(95.23±3.06)%;其誘導(dǎo)分化7天后,75%的細(xì)胞表現(xiàn)為β-TubulinⅢ陽性的神經(jīng)元樣細(xì)胞,33%的細(xì)胞則為GFAP陽性的膠質(zhì)細(xì)胞。 結(jié)論:骨髓基質(zhì)干細(xì)胞在體外培養(yǎng)條件下生長良好,經(jīng)bFGF㧏EGF誘導(dǎo)后可大量分化為神經(jīng)元樣細(xì)胞。 第二部分大鼠骨髓基質(zhì)干細(xì)胞定向誘導(dǎo)分化為神經(jīng)元樣細(xì)胞的相關(guān)分子機制研究 目的:探索大鼠骨髓基質(zhì)干細(xì)胞(bone marrow stromal cells, BMSCs)體外定向誘導(dǎo)分化為神經(jīng)元樣細(xì)胞所涉及的相關(guān)基因,并嘗試探討這些基因在BMSCs誘導(dǎo)分化為神經(jīng)元樣細(xì)胞時所起的作用。 方法:體外培養(yǎng)BMSCs至第4代,以bFGF+EGF分別誘導(dǎo)0,4,7天,常規(guī)抽提和純化RNA,采用Affymetrix Rat Expression Array 230 2.0芯片分析誘導(dǎo)前后的基因表達譜改變,找出顯著上調(diào)及下調(diào)的基因。 結(jié)果:通過對誘導(dǎo)前后的基因表達譜進行比較,總共找到97條重要表達基因,其中上調(diào)2倍以上的基因62條(其中有17條與神經(jīng)元相關(guān)),下調(diào)2倍以上的基因35條(其中有7條與神經(jīng)元相關(guān))。 結(jié)論:在BMSCs體外定向誘導(dǎo)分化為神經(jīng)元樣細(xì)胞的過程中,至少有24條重要基因參與調(diào)控,值得進一步深入研究。
[Abstract]:Part one: isolation, Culture and directional differentiation of Rat Bone Marrow Stromal Stem cells in VitroAim: to explore the suitable conditions for isolation, culture and purification of rat bone marrow stromal stem cells (BMSCs) in vitro, and to explore the feasibility of differentiation into neuron-like cells induced by BMSCs in vitro.Methods: BMSCs were isolated from adult rat bone marrow by density gradient centrifugation.The purified BMSCs was induced by basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF), and the differentiation results were identified by immunocytochemical staining.Results: the purity of BMSCs cultured in vitro was the highest (95.23 鹵3.06) after 4 passages, and 75% of the cells showed 尾 -Tubulin 鈪，

本文編號：1709763