本文選題：神經(jīng)干細(xì)胞 + 細(xì)胞　； 參考：《中國(guó)組織工程研究》2017年17期

【摘要】：背景:神經(jīng)干細(xì)胞在神經(jīng)損傷修復(fù)和退行性疾病中有廣泛的應(yīng)用前景,體外培養(yǎng)鑒定及促神經(jīng)元誘導(dǎo)分化是后續(xù)研究的基礎(chǔ)。目的:采用懸浮神經(jīng)球培養(yǎng)法分離培養(yǎng)神經(jīng)干細(xì)胞并對(duì)其鑒定,了解生物學(xué)特性。方法:采用懸浮神經(jīng)球培養(yǎng)法從C57BL/6胎鼠大腦半球分離培養(yǎng)神經(jīng)干細(xì)胞,觀察形態(tài)特征及超微結(jié)構(gòu),CCK-8法繪制生長(zhǎng)曲線,流式細(xì)胞儀檢測(cè)細(xì)胞周期,免疫熒光法檢測(cè)特異性標(biāo)志蛋白Nestin的表達(dá);用體積分?jǐn)?shù)為1%和10%的血清誘導(dǎo)分化后,免疫熒光法檢測(cè)GFAP、βⅢ-tubulin和MBP的表達(dá)。結(jié)果與結(jié)論:(1)體外培養(yǎng)得到懸浮生長(zhǎng)的神經(jīng)球,生長(zhǎng)曲線和細(xì)胞周期表明細(xì)胞增殖力強(qiáng);(2)透射電鏡觀察到神經(jīng)干細(xì)胞核漿比高,呈未分化狀態(tài);(3)Nestin免疫熒光陽(yáng)性;(4)不同體積分?jǐn)?shù)的血清誘導(dǎo)后均可分化為星形膠質(zhì)細(xì)胞、神經(jīng)元和少突膠質(zhì)細(xì)胞,體積分?jǐn)?shù)為1%血清能誘導(dǎo)分化得到更多的神經(jīng)元細(xì)胞;(5)結(jié)果表明,采用懸浮神經(jīng)球培養(yǎng)法成功分離培養(yǎng)得到了神經(jīng)干細(xì)胞,低體積分?jǐn)?shù)血清有利于神經(jīng)干細(xì)胞向神經(jīng)元分化。
[Abstract]:Background: neural stem cells (NSCs) are widely used in nerve injury repair and degenerative diseases. In vitro culture and identification of neural stem cells and induction of neuronal differentiation are the basis of further research. Objective: to isolate and identify neural stem cells (NSCs) by suspension culture. Methods: neural stem cells were isolated and cultured from the cerebral hemisphere of C57BL / 6 fetal mice by suspension ball culture method. The morphology and ultrastructure of neural stem cells were observed and the growth curve was drawn by CCK-8 method. The cell cycle was measured by flow cytometry. The expression of nestin, a specific marker protein, was detected by immunofluorescence, and the expression of GFAP, 尾 鈪，

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