Simple Derivation of Spinal Motor Neurons from ESCs/iPSCs Using Sendai Virus Vectors.
Simple Derivation of Spinal Motor Neurons from ESCs/iPSCs Using Sendai Virus Vectors.
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DOI：
10.1016/j.omtm.2016.12.007
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发表时间：
2017-03-17
期刊：
影响因子：
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通讯作者：
Inoue H
中科院分区：
文献类型：
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作者：
Goto K;Imamura K;Komatsu K;Mitani K;Aiba K;Nakatsuji N;Inoue M;Kawata A;Yamashita H;Takahashi R;Inoue H
肌萎缩侧索硬化症（amyotrophiclateralsclerosis，ALS）是一种进行性和致死性的运动神经元退行性疾病。胚胎干细胞（ESC）/诱导多能干细胞（iPSC）现在帮助我们通过疾病建模来了解ALS的病理机制。已经报道了通过添加信号传导分子将ESC/iPSC分化为MN的各种方法。然而，经典方法需要多个步骤，并且使用转录因子转导的更新的简单方法存在载体基因的基因组整合的风险。转录因子表达水平的异质性也仍然是一个问题。在这里，我们描述了一种新的方法，用于分化人类和小鼠胚胎干细胞/iPSCs成MN使用一个单一的仙台病毒载体编码三个转录因子，LIM/同源框蛋白3，神经生成素2，和胰岛-1，这是整合免费。与使用三种单独的仙台病毒载体相比，这种在第2天从人iPSC产生HB 9阳性细胞的单载体方法增加了MN与神经元的比率。此外，通过该方法从ALS患者和模型小鼠的iPSC衍生的MN显示疾病表型。这种简单的方法大大减少了生成MN所需的工作量，并为疾病建模提供了有用的工具。
Amyotrophic lateral sclerosis (ALS) is a progressive and fatal degenerative disorder of motor neurons (MNs). Embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs) now help us to understand the pathomechanisms of ALS via disease modeling. Various methods to differentiate ESCs/iPSCs into MNs by the addition of signaling molecules have been reported. However, classical methods require multiple steps, and newer simple methods using the transduction of transcription factors run the risk of genomic integration of the vector genes. Heterogeneity of the expression levels of the transcription factors also remains an issue. Here we describe a novel approach for differentiating human and mouse ESCs/iPSCs into MNs using a single Sendai virus vector encoding three transcription factors, LIM/homeobox protein 3, neurogenin 2, and islet-1, which are integration free. This single-vector method, generating HB9-positive cells on day 2 from human iPSCs, increases the ratio of MNs to neurons compared to the use of three separate Sendai virus vectors. In addition, the MNs derived via this method from iPSCs of ALS patients and model mice display disease phenotypes. This simple approach significantly reduces the efforts required to generate MNs, and it provides a useful tool for disease modeling.