Chen Chun-Yuan, Liao Wei, Lou Yuan-Lei, Li Qing, Hu Bin, Wang Yang, Deng Zhi-Feng
Graduate School of Nanchang University, Nanchang, 330006, China.
Mol Cell Biochem. 2014 Oct;395(1-2):291-8. doi: 10.1007/s11010-014-2130-3. Epub 2014 Jun 28.
Neural stem cells (NSCs) derived from induced pluripotent stem cells (iPSCs) are becoming an appealing source of cell-based therapies of brain diseases. As such, it is important to understand the molecular mechanisms that regulate the differentiation of iPSCs toward NSCs. It is well known that Notch signaling governs the retention of stem cell features and drives stem cells fate. However, further studies are required to investigate the role of Notch signaling in the NSCs differentiation of iPSCs. In this study, we successfully generated NSCs from human iPSCs using serum-free medium supplemented with retinoic acid (RA) in vitro. We then assessed changes in the expression of Notch signaling-related molecules and some miRNAs (9, 34a, 200b), which exert their regulation by targeting Notch signaling. Moreover, we used a γ-secretase inhibitor (DAPT) to disturb Notch signaling. Data revealed that the levels of the Notch signaling-related molecules decreased, whereas those miRNAs increased, during this differentiation process. Inhibition of Notch signaling accelerated the formation of the neural rosette structures and the expression of NSC and mature neurocyte marker genes. This suggests that Notch signaling negatively regulated the neuralization of human iPSCs, and that this process may be regulated by some miRNAs.
源自诱导多能干细胞（iPSC）的神经干细胞（NSC）正成为脑部疾病细胞疗法的一个有吸引力的细胞来源。因此，了解调控iPSC向NSC分化的分子机制非常重要。众所周知，Notch信号通路控制干细胞特征的维持并驱动干细胞命运。然而，需要进一步研究来探究Notch信号通路在iPSC的NSC分化中的作用。在本研究中，我们在体外使用添加了视黄酸（RA）的无血清培养基成功地从人iPSC中生成了NSC。然后，我们评估了Notch信号通路相关分子以及一些通过靶向Notch信号通路发挥调控作用的miRNA（9、34a、200b）的表达变化。此外，我们使用γ-分泌酶抑制剂（DAPT）来干扰Notch信号通路。数据显示，在这个分化过程中，Notch信号通路相关分子的水平下降，而那些miRNA的水平上升。Notch信号通路的抑制加速了神经玫瑰花结结构的形成以及NSC和成熟神经细胞标志物基因的表达。这表明Notch信号通路对人iPSC的神经化起负调控作用，并且这个过程可能受一些miRNA调控。
