【摘要】： 骨髓間充質(zhì)干細(xì)胞(BMSCs)成為視網(wǎng)膜細(xì)胞移植的理想材料,通過(guò)對(duì)bFGF刺激骨髓間充質(zhì)干細(xì)胞增殖分化后神經(jīng)干細(xì)胞和神經(jīng)細(xì)胞特異性標(biāo)志物表達(dá)的檢測(cè),探討bFGF在骨髓間充質(zhì)干細(xì)胞向視網(wǎng)膜神經(jīng)樣細(xì)胞誘導(dǎo)分化過(guò)程中的作用。 體外分離提取骨髓細(xì)胞進(jìn)行培養(yǎng),傳至第3代時(shí)經(jīng)流式細(xì)胞儀檢測(cè)細(xì)胞表面標(biāo)志物CD90、CD71、CD29表達(dá)陽(yáng)性,CD45表達(dá)陰性,免疫細(xì)胞化學(xué)方法鑒定CD71(+),CD90(+)。將BMSCs細(xì)胞分為兩組,實(shí)驗(yàn)組分別加入濃度為10ng/ml,20ng/ml,30ng/ml的bFGF,對(duì)照組不加因子。培養(yǎng)9天后,免疫細(xì)胞化學(xué)染色檢測(cè)實(shí)驗(yàn)組巢蛋白(Nestin)、神經(jīng)元特異性烯醇化酶(NSE)表達(dá)陽(yáng)性,對(duì)照組表達(dá)陰性。各組經(jīng)RT-PCR檢測(cè)均表達(dá)βⅢ-tubulin、nestin、NSE mRNA,實(shí)驗(yàn)組高于對(duì)照組,實(shí)驗(yàn)組各濃度組未見明顯差異。兩組細(xì)胞更換無(wú)血清的DMEM/F12培養(yǎng)基誘導(dǎo)2h后部分細(xì)胞成神經(jīng)樣形態(tài)改變。免疫細(xì)胞化學(xué)染色檢測(cè)兩組均表nestin、NSE;用RT-PCR方法均可檢測(cè)到βⅢ-tubulin、nestin、NSE mRNA的表達(dá),實(shí)驗(yàn)組表達(dá)高于對(duì)照組,實(shí)驗(yàn)組各濃度組未見明顯差異。 對(duì)大鼠BMSCs進(jìn)行有效的分離、培養(yǎng)可得到較均一的BMSCs。應(yīng)用無(wú)血清培養(yǎng)基誘導(dǎo)的方法可將BMSCs誘導(dǎo)分化為神經(jīng)樣細(xì)胞。bFGF對(duì)BMSCs具有促進(jìn)增殖的作用,并在向視網(wǎng)膜神經(jīng)樣細(xì)胞定向分化過(guò)程中起到促進(jìn)作用。
[Abstract]:Bone marrow mesenchymal stem cell (BMSCs) is an ideal material for retinal cell transplantation. The expression of neural stem cells and neuronal specific markers after proliferation and differentiation of bone marrow mesenchymal stem cells stimulated by bFGF was detected. To investigate the role of bFGF in the differentiation of bone marrow mesenchymal stem cells into retinal neuronal cells. Bone marrow cells were isolated and cultured in vitro. At the third passage, positive expression of CD90,CD71,CD29 and negative expression of CD45 were detected by flow cytometry. CD71 (), CD90 (). Was identified by immunocytochemistry. BMSCs cells were divided into two groups. The experimental group was treated with 10 ng / ml 20 ng / ml bFGF, control group and 30 ng / ml bFGF, control group. After 9 days of culture, the positive expression of nestin (Nestin), neuron specific enolase (NSE) was detected by immunocytochemical staining in the experimental group and negative in the control group. The expression of 尾-鈪，

本文編號(hào)：2265831