Department of Molecular and Cellular Physiology, Stanford University School of Medicine, 265 Campus Drive, Stanford, CA 94305, USA.
Neuron. 2013 Jun 5;78(5):785-98. doi: 10.1016/j.neuron.2013.05.029.
Available methods for differentiating human embryonic stem cells (ESCs) and induced pluripotent cells (iPSCs) into neurons are often cumbersome, slow, and variable. Alternatively, human fibroblasts can be directly converted into induced neuronal (iN) cells. However, with present techniques conversion is inefficient, synapse formation is limited, and only small amounts of neurons can be generated. Here, we show that human ESCs and iPSCs can be converted into functional iN cells with nearly 100% yield and purity in less than 2 weeks by forced expression of a single transcription factor. The resulting ES-iN or iPS-iN cells exhibit quantitatively reproducible properties independent of the cell line of origin, form mature pre- and postsynaptic specializations, and integrate into existing synaptic networks when transplanted into mouse brain. As illustrated by selected examples, our approach enables large-scale studies of human neurons for questions such as analyses of human diseases, examination of human-specific genes, and drug screening.
现有的将人类胚胎干细胞(ESCs)和诱导多能干细胞(iPSCs)分化为神经元的方法通常繁琐、缓慢且多变。或者,可以将人成纤维细胞直接转化为诱导性神经元(iN)细胞。然而,目前的技术转化率低,突触形成受限,只能产生少量的神经元。在这里,我们展示了通过强制表达单个转录因子,人类 ESCs 和 iPSCs 可以在不到 2 周的时间内高效地转化为功能性 iN 细胞,转化率接近 100%,纯度接近 100%。所得的 ES-iN 或 iPS-iN 细胞表现出定量重现的特性,与起始细胞系无关,形成成熟的前突触和后突触特化,并在移植到小鼠大脑时整合到现有的突触网络中。如通过选定的例子所示,我们的方法可以大规模地研究人类神经元,以解决人类疾病分析、人类特异性基因研究和药物筛选等问题。