Inhibition of Notch Signaling in Human Embryonic Stem Cell-Derived Neural Stem Cells Delays G1/S Phase Transition and Accelerates Neuronal Differentiation In Vitro and In Vivo
Inhibition of Notch Signaling in Human Embryonic Stem Cell-Derived Neural Stem Cells Delays G1/S Phase Transition and Accelerates Neuronal Differentiation In Vitro and In Vivo
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DOI：
10.1002/stem.408
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发表时间：
2010-05-01
期刊：
影响因子：
5.2
通讯作者：
Bruestle, Oliver
中科院分区：
文献类型：
--
作者：
Borghese, Lodovica;Dolezalova, Dasa;Bruestle, Oliver
人类胚胎干细胞(HESCs)和其他多能干细胞的体外可控分化为产生大量的人类神经元提供了有趣的前景，用于各种生物医学应用。与这种方法相关的一个主要瓶颈是hESC来源的神经细胞需要很长时间才能产生成熟的神经元后代。在脊椎动物发育的神经系统中，Notch信号是神经干细胞(NSC)维持的关键调节因子。在这里，我们开始探索这一信号通路是否可以被用来调节hESC来源的神经干细胞(HESNSCs)的分化。我们检测了Notch通路在hESNSCs中的表达，证明了Notch信号在自我更新培养条件下是活跃的。C-分泌酶抑制剂N-[N-(3，5-difluorophenacetyl)-L-alanyl]-S-phenylglycinet-butyl酯(DAPT)对人胚胎干细胞中Notch活性的抑制会影响Notch已知靶点的人类同源物和几种细胞周期调节因子的表达。此外，DAPT介导的Notch抑制延迟了G1/S期的转变，并使hESNSCs参与了神经发生。结合生长因子的撤除，抑制Notch信号导致明显的分化加速，从而缩短电生理活性的hESNSC来源神经元的产生时间。这一效应可用于神经细胞移植，在移植前短暂抑制Notch足以促进宿主组织中hESNSCs的神经元分化。因此，对Notch信号的干扰为在体外和体内控制人类NSC分化提供了一种工具。干细胞2010；28：955-964
The controlled in vitro differentiation of human embryonic stem cells (hESCs) and other pluripotent stem cells provides interesting prospects for generating large numbers of human neurons for a variety of biomedical applications. A major bottleneck associated with this approach is the long time required for hESC-derived neural cells to give rise to mature neuronal progeny. In the developing vertebrate nervous system, Notch signaling represents a key regulator of neural stem cell (NSC) maintenance. Here, we set out to explore whether this signaling pathway can be exploited to modulate the differentiation of hESC-derived NSCs (hESNSCs). We assessed the expression of Notch pathway components in hESNSCs and demonstrate that Notch signaling is active under self-renewing culture conditions. Inhibition of Notch activity by the c-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycinet-butyl ester (DAPT) in hESNSCs affects the expression of human homologues of known targets of Notch and of several cell cycle regulators. Furthermore, DAPT-mediated Notch inhibition delays G1/S-phase transition and commits hESNSCs to neurogenesis. Combined with growth factor withdrawal, inhibition of Notch signaling results in a marked acceleration of differentiation, thereby shortening the time required for the generation of electrophysiologically active hESNSC-derived neurons. This effect can be exploited for neural cell transplantation, where transient Notch inhibition before grafting suffices to promote the onset of neuronal differentiation of hESNSCs in the host tissue. Thus, interference with Notch signaling provides a tool for controlling human NSC differentiation both in vitro and in vivo. STEM CELLS 2010; 28: 955-964