小鼠iPS细胞体外自发性分化及神经定向分化中基因表达谱的研究

发布时间：2018-05-09 16:11

本文选题：诱导多能干细胞 + 自发性分化　；参考：《神经解剖学杂志》2017年03期

【摘要】：目的:研究小鼠诱导多能干细胞(i PS细胞)在体外自发性分化和神经定向分化过程中的基因表达谱的特性。方法:在体外将i PS细胞分别进行形成类胚胎小体后自发性分化及成骨蛋白抑制剂Noggin诱导神经定向分化后,通过实时荧光定量PCR(q PCR)测定在分化过程中i PS细胞分化相关基因表达的变化。结果:i PS细胞形成类胚胎小体后胚胎外胚层标志基因(GFAP,Map2和Tu J1)及内胚层标记基因(Foxa2,GATA4和Sox17)的表达随分化而迅速增加,但中胚层标记基因(Bmp4,BRA,FGF5)表达水平变化不明显。在神经定向分化过程中,i PS细胞的神经标志物基因(Map2,Neu N,Tu J1和Sox1)及线粒体相关基因(12s,ND3,ND5,ND6、Cytb和Cox1)的表达都逐渐增加,且两者具有相同的增长趋势。结论:在自发性分化过程中小鼠i PS细胞具有自发向外胚层和内胚层分化的趋势;在神经定向分化过程中,i PS细胞的线粒体相关基因表达随着分化表达逐渐增加,并与神经细胞标记基因具有正相关性,表明线粒体的功能在i PS细胞神经定向分化中发挥重要作用。
[Abstract]:Aim: to study the characteristics of gene expression profile of mouse induced pluripotent stem cell / PS cells during spontaneous and neuronal differentiation in vitro. Methods: in vitro, I PS cells were divided into embryoid bodies after spontaneous differentiation and osteogenic protein inhibitor Noggin induced neuronal directional differentiation. The expression of differentiation related genes in iPS cells was measured by real-time fluorescence quantitative PCR(q. Results the expression of embryonic ectodermal marker genes (GFAPMap2 and Tu J1) and endodermal marker genes (Foxa2 (GATA4) and Sox17) increased rapidly with differentiation, but the expression level of mesodermal marker gene (Bmp4, BRAFGF5) did not change significantly. During the process of neuronal directional differentiation, the expression of neuronal marker gene (map2Neu Nu Tu J1 and Sox1) and mitochondrial associated gene (12sNd3Nd5Nd5Nd6, Cytb and Cox1) in PS cells increased gradually, and both of them had the same increasing trend. Conclusion: the spontaneous differentiation of mouse iPS cells into ectoderm and endoderm during spontaneous differentiation and the expression of mitochondria related genes in iPS cells increased with the differentiation. There is a positive correlation between mitochondria and neuronal marker gene, which indicates that mitochondrial function plays an important role in the neuronal differentiation of iPS cells.
【作者单位】：第四军医大学西京医院神经内科;广西医科大学第一附属医院神经内科;挪威科技大学肿瘤研究及分子医学部;
【基金】：国家自然科学基金(81571223)
【分类号】：R741

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