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bFGF对骨髓间充质干细胞向视网膜神经样细胞增殖分化的影响

发布时间:2018-10-12 10:49  文章来源:笔耕文化传播
【摘要】: 骨髓间充质干细胞(BMSCs)成为视网膜细胞移植的理想材料,通过对bFGF刺激骨髓间充质干细胞增殖分化后神经干细胞和神经细胞特异性标志物表达的检测,探讨bFGF在骨髓间充质干细胞向视网膜神经样细胞诱导分化过程中的作用。 体外分离提取骨髓细胞进行培养,传至第3代时经流式细胞仪检测细胞表面标志物CD90、CD71、CD29表达阳性,CD45表达阴性,免疫细胞化学方法鉴定CD71(+),CD90(+)。将BMSCs细胞分为两组,实验组分别加入浓度为10ng/ml,20ng/ml,30ng/ml的bFGF,对照组不加因子。培养9天后,免疫细胞化学染色检测实验组巢蛋白(Nestin)、神经元特异性烯醇化酶(NSE)表达阳性,对照组表达阴性。各组经RT-PCR检测均表达βⅢ-tubulin、nestin、NSE mRNA,实验组高于对照组,实验组各浓度组未见明显差异。两组细胞更换无血清的DMEM/F12培养基诱导2h后部分细胞成神经样形态改变。免疫细胞化学染色检测两组均表nestin、NSE;用RT-PCR方法均可检测到βⅢ-tubulin、nestin、NSE mRNA的表达,实验组表达高于对照组,实验组各浓度组未见明显差异。 对大鼠BMSCs进行有效的分离、培养可得到较均一的BMSCs。应用无血清培养基诱导的方法可将BMSCs诱导分化为神经样细胞。bFGF对BMSCs具有促进增殖的作用,并在向视网膜神经样细胞定向分化过程中起到促进作用。
[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,

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