论文部分内容阅读
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。
OBJECTIVE: To study the gene expression profile of mouse induced pluripotent stem cells (i PS cells) in vitro during spontaneous differentiation and neural differentiation. Methods: iPS cells were induced to differentiate spontaneously after the formation of embryo-like bodies and neuro-differentiation induced by osteoblast inhibitor Noggin in vitro. Differentiation of i PS cells was detected by real-time fluorescence quantitative PCR (qPCR) Related gene expression changes. Results: The expressions of embryonic ectoderm markers (GFAP, Map2 and Tu J1) and endodermal marker genes (Foxa2, GATA4 and Sox17) in iPS cell-like embryo bodies increased rapidly with differentiation, but mesodermal marker gene Bmp4, BRA, FGF5) expression did not change significantly. The neuronal marker genes (Map2, Neu N, Tu J1 and Sox1) and mitochondrial related genes (12s, ND3, ND5, ND6, Cytb and Cox1) in iPS cells gradually increased in the process of neuro-directional differentiation Both have the same growth trend. CONCLUSIONS: The mouse PSSCs have the tendency of spontaneous differentiation to ectoderm and endoderm during spontaneous differentiation. In the process of neural differentiation, the expression of mitochondria-related genes in i PS cells gradually increases with the differentiation and expression, The marker gene has a positive correlation, suggesting that mitochondrial function plays an important role in the i nterminal neuronal differentiation of i PS cells.