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将人类诱导多能干细胞(iPSCs)分化为前脑神经祖细胞和成熟神经元的有效模型。

Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) into an Effective Model of Forebrain Neural Progenitor Cells and Mature Neurons.

作者信息

Bell Scott, Hettige Nuwan C, Silveira Heika, Peng Huashan, Wu Hanrong, Jefri Malvin, Antonyan Lilit, Zhang Ying, Zhang Xin, Ernst Carl

机构信息

Psychiatric Genetics Group, McGill University and Douglas Hospital Research Institute, Department of Psychiatry, Verdun, Montreal, QC H4H 1R3, Canada.

出版信息

Bio Protoc. 2019 Mar 5;9(5):e3188. doi: 10.21769/BioProtoc.3188.

Abstract

Induced Pluripotent Stem Cells (iPSCs) are pluripotent stem cells that can be generated from somatic cells, and provide a way to model the development of neural tissues . One particularly interesting application of iPSCs is the development of neurons analogous to those found in the human forebrain. Forebrain neurons play a central role in cognition and sensory processing, and deficits in forebrain neuronal activity contributes to a host of conditions, including epilepsy, Alzheimer's disease, and schizophrenia. Here, we present our protocol for differentiating iPSCs into forebrain neural progenitor cells (NPCs) and neurons, whereby neural rosettes are generated from stem cells without dissociation and NPCs purified from rosettes based on their adhesion, resulting in a more rapid generation of pure NPC cultures. Neural progenitor cells can be maintained as long-term cultures, or differentiated into forebrain neurons. This protocol provides a simplified and fast methodology of generating forebrain NPCs and neurons, and enables researchers to generate effective models to study forebrain disease and neurodevelopment. This protocol can also be easily adapted to generate other neural lineages.

摘要

诱导多能干细胞(iPSC)是一类可从体细胞生成的多能干细胞,为模拟神经组织发育提供了一种方法。iPSC的一个特别有趣的应用是培育出类似于人类前脑神经元的神经元。前脑神经元在认知和感觉处理中起着核心作用,前脑神经元活动的缺陷会导致一系列病症,包括癫痫、阿尔茨海默病和精神分裂症。在此,我们展示了将iPSC分化为前脑神经祖细胞(NPC)和神经元的方案,即从干细胞生成神经玫瑰花结而不进行解离,并根据其黏附性从玫瑰花结中纯化NPC,从而更快地生成纯NPC培养物。神经祖细胞可作为长期培养物进行维持,或分化为前脑神经元。该方案提供了一种生成前脑NPC和神经元的简化且快速的方法,使研究人员能够生成有效的模型来研究前脑疾病和神经发育。该方案也可轻松调整以生成其他神经谱系。

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