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Transcription Factor–MediatedDifferentiation of Human iPSCsinto Neurons

机译:转录因子 - 人IPSCsinto神经元的介导

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摘要

Accurate modeling of human neuronal cell biology has been a long-standingchallenge. However, methods to differentiate human induced pluripotent stemcells (iPSCs) to neurons have recently provided experimentally tractable cellmodels. Numerous methods that use small molecules to direct iPSCs intoneuronal lineages have arisen in recent years. Unfortunately, these methodsentail numerous challenges, including poor efficiency, variable cell type heterogeneity,and lengthy, expensive differentiation procedures. We recently developeda new method to generate stable transgenic lines of human iPSCs withdoxycycline-inducible transcription factors at safe-harbor loci. Using a simpletwo-step protocol, these lines can be inducibly differentiated into either cortical(i3Neurons) or lower motor neurons (i3LMN) in a rapid, efficient, and scalablemanner (Wang et al., 2017). In this manuscript, we describe a set of protocolsto assist investigators in the culture and genetic engineering of iPSC lines toenable transcription factor–mediated differentiation of iPSCs into i3Neurons ori3LMNs, and we present neuronal culture conditions for various experimentalapplications.
机译:准确的人类神经元细胞生物学建模一直是一个长姿态的。然而,最近提供了将人类诱导的多能干细胞(IPSC)分化为神经元的方法,最近提供了实验易易腐败的细胞巨。近年来,利用小分子使用小分子直接IPSC的方法。不幸的是,这些方法南部众多挑战,包括效率差,可变细胞类型异质性,以及冗长,昂贵的分化程序。我们最近发育了在安全 - 港口基因座的氧化环素 - 诱导转录因子中产生稳定的转基因患者的新方法。使用简单的行业协议,可以诱导这些线条在快速,高效和ScalableManner(Wang等,2017)中诱导皮质(I3neurons)或下电机神经元(I3LMN)。在本手稿中,我们描述了一套协议助理调查人员在IPSC系列的培养和基因工程中辅助调查人员IPSCS转录因子介导的IPSCS进入IPNURONS ORI3LMNS,并且我们为各种实验性申请呈现神经元培养条件。

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