【摘要】： 目的 研究原代人臍帶間充質干細胞(UCMSCs)的生物學特性及其體外向神經(jīng)細胞誘導分化的條件,為組織化人工神經(jīng)的制備提供干細胞資源。 方法 本實驗中我們通過在無菌條件下取正常足月剖腹產新生兒的臍帶,利用組織塊培養(yǎng)法獲得貼壁細胞并進行傳代培養(yǎng)和純化。取第2代貼壁細胞,免疫組化測定細胞表面CD44、CD105、CD106、CD34、CD45表達情況。取生長狀態(tài)良好的UCMSCs進行細胞爬片,待細胞生長至60%-70%融合時分為四組進行誘導分化。四組細胞分別加入單純培養(yǎng)液空白對照組(A組)、單純生長因子誘導組(B組)、單純黃芪誘導組(C組)和黃芪聯(lián)合生長因子誘導液(D組),倒置顯微鏡下觀察其形態(tài)并檢測其表面神經(jīng)細胞的標志物如NSE、NF和GFAP的陽性表達率,將結果進行統(tǒng)計學分析。 結果 1.原代培養(yǎng)的組織塊在5-6d后可見新生貼壁的細胞,形態(tài)類似成纖維細胞,一周后快速增殖,成漩渦狀聚集,在生長的過程中呈現(xiàn)間充質干細胞的形態(tài)特點,免疫組化檢測表面標記CD44、CD105強陽性,CD106弱陽性,CD34、CD45陰性,檢測結果和UCMSCs的生物學特征符合。 2.單純bFGF、單純黃芪和黃芪聯(lián)合生長因子都可以將細胞誘導向神經(jīng)樣細胞分化,但是通過誘導后細胞表面神經(jīng)標志物的測定,黃芪聯(lián)合生長因子誘導組的細胞NSE、NF和GFAP的陽性表達率也更高,經(jīng)統(tǒng)計學分析,兩者的差異明顯,具有統(tǒng)計學意義。 結論 1.中藥黃芪聯(lián)合生長因子可明顯促進人UCMSCs分化為神經(jīng)樣細胞,不僅細胞形態(tài)發(fā)生明顯變化,而且從分子生物學水平表現(xiàn)出神經(jīng)細胞特征。 2.與單純生長因子誘導組(B組)和單純黃芪誘導組(C組)相比,黃芪聯(lián)合生長因子誘導(D組)誘導后的人UCMSCs的NSE、NF和GFAP的陽性表達率也更高,經(jīng)統(tǒng)計學分析,兩者的差異明顯,具有統(tǒng)計學意義。
[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，

本文編號：2474923