Transcription Factor-Mediated Differentiation of Human iPSCs into Neurons.
Transcription Factor-Mediated Differentiation of Human iPSCs into Neurons.
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DOI：
10.1002/cpcb.51
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发表时间：
2018-06
影响因子：
--
通讯作者：
Ward ME
中科院分区：
文献类型：
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作者：
Fernandopulle MS;Prestil R;Grunseich C;Wang C;Gan L;Ward ME
人类神经元细胞生物学的精确建模一直是一个长期的挑战。然而，将人类诱导多能干细胞（iPSCs）分化为神经元的方法最近提供了实验可处理的细胞模型。近年来出现了许多使用小分子引导iPSCs进入神经元谱系的方法。不幸的是，这些方法带来了许多挑战，包括效率低，细胞类型同质性可变，以及冗长，复杂和昂贵的分化过程。我们最近开发了一种新的方法，在安全港位点产生具有强力霉素诱导的转录因子的稳定的人多能干细胞转基因系。使用简单的两步方案，这些细胞系可以以快速、有效和可扩展的方式诱导分化为皮层（i3neuron）或下层运动神经元（i3LMN）。在这篇手稿中，我们描述了一套方案，以协助研究人员在培养和基因工程的iPSC系，使转录因子介导的iPSC分化成i3neuron或i3lmn，我们提出了各种实验应用的神经元培养条件。
Accurate modeling of human neuronal cell biology has been a long-standing challenge. However, methods to differentiate human induced pluripotent stem cells (iPSCs) to neurons have recently provided experimentally tractable cell models. Numerous methods that use small molecules to direct iPSCs into neuronal lineages have arisen in recent years. Unfortunately, these methods entail numerous challenges, including poor efficiency, variable cell-type homogeneity, and lengthy, complicated, and expensive differentiation procedures. We recently developed a new method to generate stable transgenic lines of human iPSCs with doxycycline-inducible transcription factors at safe-harbor loci. Using a simple two-step protocol, these lines can be inducibly differentiated into either cortical (i3Neurons) or lower motor neurons (i3LMN) in a rapid, efficient, and scalable manner. In this manuscript, we describe a set of protocols to assist investigators in the culture and genetic engineering of iPSC lines to enable transcription-factor mediated differentiation of iPSCs into i3Neurons or i3LMNs, and we present neuronal culture conditions for various experimental applications.