目的探討丹參誘導(dǎo)骨隨間充質(zhì)干細(xì)胞(Mesenchymal stem cells, MSC)分化為神經(jīng)樣細(xì)胞優(yōu)化方案可行性及分化后的神經(jīng)樣細(xì)胞是否具有神經(jīng)生理功能。 方法取4～5代狀態(tài)好的MSC加入含10%胎牛血清(Fetal bovine serum, FBS)及10 ng/ml的堿性成纖維生長因子(basic Fibroblast Growth Factor, bFGF)的培養(yǎng)基預(yù)誘導(dǎo)24 h,更換為含60 mg/L丹參液和10 ng/ml bFGF的無血清培養(yǎng)基(誘導(dǎo)液)誘導(dǎo)2 h,加FBS(工作濃度為10%)進(jìn)行去分化,約24 h后再次更換為誘導(dǎo)液誘導(dǎo)3 h,如此重復(fù)4⁵次,且每次誘導(dǎo)時(shí)間較前延長1 h,誘導(dǎo)后分別進(jìn)行免疫熒光細(xì)胞化學(xué),細(xì)胞膜電位(Membrane potential, MP)、細(xì)胞內(nèi)鈣流以及神經(jīng)突觸循環(huán)功能檢測(cè)。以50 mmol/L的KCl作為誘發(fā)動(dòng)作電位刺激因子,采用激光共聚焦顯微鏡(Laser-scanning confocal microscope, LSCM)對(duì)DiBAC4(3)、Fluo-3/AM、SynaptoRed C-2進(jìn)行...

【文章頁數(shù)】：121 頁

【學(xué)位級(jí)別】：博士

【部分圖文】：

圖1丹參誘導(dǎo)MSC分化為神經(jīng)樣細(xì)胞優(yōu)化方案流程圖

圖1丹參誘導(dǎo)MSC分化為神經(jīng)樣細(xì)胞優(yōu)化方案流程圖Fig1TheflowchartoftheprocedurethatMSCwasinducetodifferentiatedintoneuron-likecellsbysalviamilto....

圖2培養(yǎng)2天的原代MSC，細(xì)胞呈短梭形，小集落分布（倒置相差顯微鏡,100×）

圖1丹參誘導(dǎo)MSC分化為神經(jīng)樣細(xì)胞優(yōu)化方案流程圖Fig1TheflowchartoftheprocedurethatMSCwasinducetodifferentiatedintoneuron-likecellsbysalviamilto....

圖3培養(yǎng)7天的原代MSC，細(xì)胞呈成纖維樣，漩渦狀排列.

圖1丹參誘導(dǎo)MSC分化為神經(jīng)樣細(xì)胞優(yōu)化方案流程圖Fig1TheflowchartoftheprocedurethatMSCwasinducetodifferentiatedintoneuron-likecellsbysalviamilto....

圖4第4代的MSCs，細(xì)胞呈成纖維樣，漩渦列.（倒置相差顯微鏡，100×）

圖1丹參誘導(dǎo)MSC分化為神經(jīng)樣細(xì)胞優(yōu)化方案流程圖Fig1TheflowchartoftheprocedurethatMSCwasinducetodifferentiatedintoneuron-likecellsbysalviamilto....

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