A Step-by-Step Refined Strategy for Highly Efficient Generation of Neural Progenitors and Motor Neurons from Human Pluripotent Stem Cells.
A Step-by-Step Refined Strategy for Highly Efficient Generation of Neural Progenitors and Motor Neurons from Human Pluripotent Stem Cells.
复制标题
从人类多能干细胞高效产生神经前体细胞和运动神经元的循序渐进的改进策略。
作者：
Ren J;Li C;Zhang M;Wang H;Xie Y;Tang Y
人类神经元，特别是运动神经元（MN）的有限获取是研究神经生物学和神经疾病的主要挑战。人多能干细胞（human pluripotent stem cells，hPSC）可被诱导为神经祖细胞（neural progenitor cells，NPC）并进一步分化为多种神经亚型，为研究神经发育、细胞治疗、疾病建模和药物筛选提供了良好的细胞来源。因此，重要的是建立强大的和高效的神经分化方法。人们付出了巨大的努力来剖析神经承诺期间的关键信号，并相应地建立可靠的分化方案。在这项研究中，我们改进了一种分步策略，使hPSC在短短18天内快速分化为NPC，结合了粘附和神经球漂浮方法，以及通过引入约21天的转录因子组从NPC高效生成MN（约90%）。该策略分别利用并比较了视黄酸（RA）诱导和双SMAD通路抑制对神经诱导的影响。这两种方法都可以产生高效和完整的可识别NPC，但具有不同的区域身份。考虑到所产生的NPC可以分化成大多数兴奋性和抑制性神经元，但几乎不能分化成MN，因此我们通过过表达精细的转录因子组，特别是通过添加人SOX 11，同时改善一系列分化条件以产生成熟的MN，从而进一步将NPC分化成MN，用于运动神经元疾病的良好建模。因此，我们完善了一个详细的分步策略，用于诱导hPSC向长期可移植的NPC，并进一步指定基于NPC平台的MN。
Limited access to human neurons, especially motor neurons (MNs), was a major challenge for studying neurobiology and neurological diseases. Human pluripotent stem cells (hPSCs) could be induced as neural progenitor cells (NPCs) and further multiple neural subtypes, which provide excellent cellular sources for studying neural development, cell therapy, disease modeling and drug screening. It is thus important to establish robust and highly efficient methods of neural differentiation. Enormous efforts have been dedicated to dissecting key signalings during neural commitment and accordingly establishing reliable differentiation protocols. In this study, we refined a step-by-step strategy for rapid differentiation of hPSCs towards NPCs within merely 18 days, combining the adherent and neurosphere-floating methods, as well as highly efficient generation (~90%) of MNs from NPCs by introducing refined sets of transcription factors for around 21 days. This strategy made use of, and compared, retinoic acid (RA) induction and dual-SMAD pathway inhibition, respectively, for neural induction. Both methods could give rise to highly efficient and complete generation of preservable NPCs, but with different regional identities. Given that the generated NPCs can be differentiated into the majority of excitatory and inhibitory neurons, but hardly MNs, we thus further differentiate NPCs towards MNs by overexpressing refined sets of transcription factors, especially by adding human SOX11, whilst improving a series of differentiation conditions to yield mature MNs for good modeling of motor neuron diseases. We thus refined a detailed step-by-step strategy for inducing hPSCs towards long-term preservable NPCs, and further specified MNs based on the NPC platform.
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影响因子：
16.6
作者：
Du, Zhong-Wei;Chen, Hong;Liu, Huisheng;Lu, Jianfeng;Qian, Kun;Huang, CindyTzu-Ling;Zhong, Xiaofen;Fan, Frank;Zhang, Su-Chun
通讯作者：
Zhang, Su-Chun
影响因子：
4.8
作者：
Kozhich, O. A.;Hamilton, R. S.;Mallon, B. S.
通讯作者：
Mallon, B. S.
影响因子：
10.5
作者：
Bergsland, Maria;Werme, Martin;Muhr, Jonas
通讯作者：
Muhr, Jonas
DOI：
10.1073/pnas.0910012107
发表时间：
2010-03-02
影响因子：
11.1
作者：
Hu, Bao-Yang;Weick, Jason P.;Zhang, Su-Chun
通讯作者：
Zhang, Su-Chun
影响因子：
46.9
作者：
Chambers, Stuart M.;Fasano, Christopher A.;Papapetrou, Eirini P.;Tomishima, Mark;Sadelain, Michel;Studer, Lorenz
通讯作者：
Studer, Lorenz