Highly Expandable Human iPS Cell-Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High-Throughput Screening.
Highly Expandable Human iPS Cell-Derived Neural Progenitor Cells (NPC) and Neurons for Central Nervous System Disease Modeling and High-Throughput Screening.
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DOI：
10.1002/cphg.33
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发表时间：
2017-01-11
影响因子：
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通讯作者：
Haggarty, Stephen J
中科院分区：
文献类型：
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作者：
Cheng, Chialin;Fass, Daniel M;Haggarty, Stephen J
将人类体细胞重编程为诱导多能干 (iPS) 细胞，极大地扩展了可用于研究中枢神经系统 (CNS) 疾病的分子和细胞机制的研究工具。要实现 iPS 细胞技术在识别新型治疗靶点和高通量药物筛选方面的前景，需要实施大规模生产特定 CNS 细胞类型的方法。在这里，我们描述了一种使用多维荧光激活细胞分选 (FACS) 来纯化由细胞表面标记物定义的 NPC 生成稳定、高度可扩展的 iPS 细胞衍生的 CNS 神经祖细胞 (NPC) 的协议。此外，我们描述了一种快速、有效且可重复的方法，通过前神经转录因子 Neurogenin 2 (iNgn2-NPC) 的诱导表达，从这些 NPC 中产生兴奋性皮质样神经元。最后，我们描述了使用 iNgn2-NPC 通过高内涵、单细胞水平自动显微镜检测来探测人类神经可塑性和中枢神经系统疾病的机制的方法。 © 2017 John Wiley & Sons, Inc. 版权所有
Reprogramming of human somatic cells into induced pluripotent stem (iPS) cells has greatly expanded the set of research tools available to investigate the molecular and cellular mechanisms underlying central nervous system (CNS) disorders. Realizing the promise of iPS cell technology for the identification of novel therapeutic targets and for high-throughput drug screening requires implementation of methods for the large-scale production of defined CNS cell types. Here we describe a protocol for generating stable, highly expandable, iPS cell-derived CNS neural progenitor cells (NPC) using multi-dimensional fluorescence activated cell sorting (FACS) to purify NPC defined by cell surface markers. In addition, we describe a rapid, efficient, and reproducible method for generating excitatory cortical-like neurons from these NPC through inducible expression of the pro-neural transcription factor Neurogenin 2 (iNgn2-NPC). Finally, we describe methodology for the use of iNgn2-NPC for probing human neuroplasticity and mechanisms underlying CNS disorders using high-content, single-cell-level automated microscopy assays. © 2017 by John Wiley & Sons, Inc.