【摘要】： 骨髓間充質(zhì)干細胞(BMSCs)成為視網(wǎng)膜細胞移植的理想材料,通過對bFGF刺激骨髓間充質(zhì)干細胞增殖分化后神經(jīng)干細胞和神經(jīng)細胞特異性標志物表達的檢測,探討bFGF在骨髓間充質(zhì)干細胞向視網(wǎng)膜神經(jīng)樣細胞誘導(dǎo)分化過程中的作用。 體外分離提取骨髓細胞進行培養(yǎng),傳至第3代時經(jīng)流式細胞儀檢測細胞表面標志物CD90、CD71、CD29表達陽性,CD45表達陰性,免疫細胞化學(xué)方法鑒定CD71(+),CD90(+)。將BMSCs細胞分為兩組,實驗組分別加入濃度為10ng/ml,20ng/ml,30ng/ml的bFGF,對照組不加因子。培養(yǎng)9天后,免疫細胞化學(xué)染色檢測實驗組巢蛋白(Nestin)、神經(jīng)元特異性烯醇化酶(NSE)表達陽性,對照組表達陰性。各組經(jīng)RT-PCR檢測均表達βⅢ-tubulin、nestin、NSE mRNA,實驗組高于對照組,實驗組各濃度組未見明顯差異。兩組細胞更換無血清的DMEM/F12培養(yǎng)基誘導(dǎo)2h后部分細胞成神經(jīng)樣形態(tài)改變。免疫細胞化學(xué)染色檢測兩組均表nestin、NSE;用RT-PCR方法均可檢測到βⅢ-tubulin、nestin、NSE mRNA的表達,實驗組表達高于對照組,實驗組各濃度組未見明顯差異。 對大鼠BMSCs進行有效的分離、培養(yǎng)可得到較均一的BMSCs。應(yīng)用無血清培養(yǎng)基誘導(dǎo)的方法可將BMSCs誘導(dǎo)分化為神經(jīng)樣細胞。bFGF對BMSCs具有促進增殖的作用,并在向視網(wǎng)膜神經(jīng)樣細胞定向分化過程中起到促進作用。
[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：2265831