Generation of region-specific and high-purity neurons from human feeder-free iPSCs
Generation of region-specific and high-purity neurons from human feeder-free iPSCs
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DOI：
10.1016/j.neulet.2021.135676
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发表时间：
2021-02-03
影响因子：
2.5
通讯作者：
Okano, Hideyuki
中科院分区：
文献类型：
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作者：
Sato, Tsukika;Imaizumi, Kent;Okano, Hideyuki
人类诱导多能干细胞（iPSCs）在阐明神经/精神疾病的分子发病机制方面具有巨大潜力。特别是，神经/精神疾病通常表现出大脑区域特异性症状，并且已经建立了从iPSC产生区域特异性神经细胞的技术，用于体外神经/精神疾病表型的详细建模。另一方面，在没有饲养细胞的情况下培养人iPSC的最新进展已经实现了高效和可重复的神经诱导。然而，传统的区域控制技术主要是基于饲养层上iPSC开发的，并且这些方法难以应用于无饲养层（ff）iPSC培养。在这项研究中，我们建立了一种新的培养系统，从人类ff-iPSCs中产生区域特异性神经细胞。该系统是无饲养层iPSC培养的最佳优化方法，并产生具有高纯度和功能性的特定神经元亚型，包括前脑皮质神经元、前脑中间神经元、中脑多巴胺能神经元和脊髓运动神经元。此外，在我们的培养系统中再现了前脑皮层神经元层规范的时间模式，这使得能够产生层特异性皮层神经元。在本培养系统中，通过使用多电极阵列和钙成像来证明神经元活性。总的来说，我们的基于ff-iPSC的培养系统将为建模各种类型的神经/精神疾病表型提供理想的平台。
Human induced pluripotent stem cells (iPSCs) have great potential to elucidate the molecular pathogenesis of neurological/psychiatric diseases. In particular, neurological/psychiatric diseases often display brain region-specific symptoms, and the technology for generating region-specific neural cells from iPSCs has been established for detailed modeling of neurological/psychiatric disease phenotypes in vitro. On the other hand, recent advances in culturing human iPSCs without feeder cells have enabled highly efficient and reproducible neural induction. However, conventional regional control technologies have mainly been developed based on on-feeder iPSCs, and these methods are difficult to apply to feeder-free (ff) iPSC cultures. In this study, we established a novel culture system to generate region-specific neural cells from human ff-iPSCs. This system is the best optimized approach for feeder-free iPSC culture and generates specific neuronal subtypes with high purity and functionality, including forebrain cortical neurons, forebrain interneurons, midbrain dopaminergic neurons, and spinal motor neurons. In addition, the temporal patterning of cortical neuron layer specification in the forebrain was reproduced in our culture system, which enables the generation of layer-specific cortical neurons. Neuronal activity was demonstrated in the present culture system by using multiple electrode array and calcium imaging. Collectively, our ff-iPSC-based culture system would provide a desirable platform for modeling various types of neurological/psychiatric disease phenotypes.