【摘要】： 目的 研究原代人臍帶間充質(zhì)干細(xì)胞(UCMSCs)的生物學(xué)特性及其體外向神經(jīng)細(xì)胞誘導(dǎo)分化的條件,為組織化人工神經(jīng)的制備提供干細(xì)胞資源。 方法 本實(shí)驗(yàn)中我們通過(guò)在無(wú)菌條件下取正常足月剖腹產(chǎn)新生兒的臍帶,利用組織塊培養(yǎng)法獲得貼壁細(xì)胞并進(jìn)行傳代培養(yǎng)和純化。取第2代貼壁細(xì)胞,免疫組化測(cè)定細(xì)胞表面CD44、CD105、CD106、CD34、CD45表達(dá)情況。取生長(zhǎng)狀態(tài)良好的UCMSCs進(jìn)行細(xì)胞爬片,待細(xì)胞生長(zhǎng)至60%-70%融合時(shí)分為四組進(jìn)行誘導(dǎo)分化。四組細(xì)胞分別加入單純培養(yǎng)液空白對(duì)照組(A組)、單純生長(zhǎng)因子誘導(dǎo)組(B組)、單純黃芪誘導(dǎo)組(C組)和黃芪聯(lián)合生長(zhǎng)因子誘導(dǎo)液(D組),倒置顯微鏡下觀察其形態(tài)并檢測(cè)其表面神經(jīng)細(xì)胞的標(biāo)志物如NSE、NF和GFAP的陽(yáng)性表達(dá)率,將結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果 1.原代培養(yǎng)的組織塊在5-6d后可見(jiàn)新生貼壁的細(xì)胞,形態(tài)類似成纖維細(xì)胞,一周后快速增殖,成漩渦狀聚集,在生長(zhǎng)的過(guò)程中呈現(xiàn)間充質(zhì)干細(xì)胞的形態(tài)特點(diǎn),免疫組化檢測(cè)表面標(biāo)記CD44、CD105強(qiáng)陽(yáng)性,CD106弱陽(yáng)性,CD34、CD45陰性,檢測(cè)結(jié)果和UCMSCs的生物學(xué)特征符合。 2.單純bFGF、單純黃芪和黃芪聯(lián)合生長(zhǎng)因子都可以將細(xì)胞誘導(dǎo)向神經(jīng)樣細(xì)胞分化,但是通過(guò)誘導(dǎo)后細(xì)胞表面神經(jīng)標(biāo)志物的測(cè)定,黃芪聯(lián)合生長(zhǎng)因子誘導(dǎo)組的細(xì)胞NSE、NF和GFAP的陽(yáng)性表達(dá)率也更高,經(jīng)統(tǒng)計(jì)學(xué)分析,兩者的差異明顯,具有統(tǒng)計(jì)學(xué)意義。 結(jié)論 1.中藥黃芪聯(lián)合生長(zhǎng)因子可明顯促進(jìn)人UCMSCs分化為神經(jīng)樣細(xì)胞,不僅細(xì)胞形態(tài)發(fā)生明顯變化,而且從分子生物學(xué)水平表現(xiàn)出神經(jīng)細(xì)胞特征。 2.與單純生長(zhǎng)因子誘導(dǎo)組(B組)和單純黃芪誘導(dǎo)組(C組)相比,黃芪聯(lián)合生長(zhǎng)因子誘導(dǎo)(D組)誘導(dǎo)后的人UCMSCs的NSE、NF和GFAP的陽(yáng)性表達(dá)率也更高,經(jīng)統(tǒng)計(jì)學(xué)分析,兩者的差異明顯,具有統(tǒng)計(jì)學(xué)意義。
[Abstract]:Objective to study the biological characteristics of primary human umbilical cord mesenchymal stem cells (UCMSCs) and the conditions for inducing differentiation into nerve cells in vitro, so as to provide stem cell resources for the preparation of organized artificial nerve cells. Methods in this experiment, the umbilical cord of normal full-term caesarean section was taken under aseptic condition, and the adherent cells were obtained by tissue culture and subcultured and purified. The expression of CD44,CD105,CD106,CD34,CD45 on the surface of adherent cells was detected by immunohistochemistry. UCMSCs, which grew well, was divided into four groups to induce differentiation when the cells grew to 60% 鈮，

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