Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
Neurosci Lett. 2021 Feb 16;746:135676. doi: 10.1016/j.neulet.2021.135676. Epub 2021 Jan 28.
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.
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的潜力。特别是，神经/精神疾病通常表现出特定脑区的症状，并且已经建立了从 iPSCs 产生特定脑区神经细胞的技术，以便在体外对神经/精神疾病表型进行详细建模。另一方面，最近在无饲养细胞的情况下培养人类 iPSCs 的技术已经实现了高效且可重复的神经诱导。然而，传统的区域控制技术主要是基于有饲养细胞的 iPSCs 开发的，这些方法难以应用于无饲养（ff）iPSC 培养。在这项研究中，我们建立了一种从人 ff-iPSC 中生成特定脑区神经细胞的新型培养系统。该系统是最佳优化的无饲养细胞 iPSC 培养方法，可生成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，我们的培养系统重现了前脑皮质神经元层特化的时间模式，从而能够产生具有特定皮层神经元层的细胞。通过使用多电极阵列和钙成像，在本培养系统中证明了神经元的活性。总之，我们基于 ff-iPSC 的培养系统将为模拟各种类型的神经/精神疾病表型提供一个理想的平台。
