Modeling of early neural development in vitro by direct neurosphere formation culture of chimpanzee induced pluripotent stem cells
Modeling of early neural development in vitro by direct neurosphere formation culture of chimpanzee induced pluripotent stem cells
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DOI：
10.1016/j.scr.2020.101749
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发表时间：
2020-04-01
影响因子：
1.2
通讯作者：
Imamura, Masanori
中科院分区：
文献类型：
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作者：
Kitajima, Ryunosuke;Nakai, Risako;Imamura, Masanori
我们的近亲黑猩猩（类人猿）的进化发育生物学对于理解人类特征的起源至关重要。然而，由于技术和伦理的限制，很难获得黑猩猩体内的发育事件。诱导多能干细胞（iPSCs）克服了体内研究的局限性，为研究发育事件提供了另一种体外模型系统。在这里，我们从成人皮肤成纤维细胞中生成了黑猩猩的iPSCs，并在体外分化培养条件下重建了早期神经发育。黑猩猩iPSCs的建立使用了简单的方法，即携带人类重编程因子的质粒载体脂质转染，并在全面的无饲料培养中维持。最终，直接神经球形成培养诱导了从黑猩猩多能干细胞中快速有效的神经干细胞分化。神经球形成的时间过程分析表明，基因表达谱和发育潜能在早期神经发育过程中发生了变化，从外表皮细胞到放射状胶质细胞。我们的iPSC培养系统是研究黑猩猩早期神经发育的分子和细胞基础以及更好地理解人类大脑进化的有力工具。
Evolutionary developmental biology of our closest living relative, the chimpanzee (Pan troglodytes), is essential for understanding the origin of human traits. However, it is difficult to access developmental events in the chimpanzee in vivo because of technical and ethical restrictions. Induced pluripotent stem cells (iPSCs) offer an alternative in vitro model system to investigate developmental events by overcoming the limitations of in vivo study. Here, we generated chimpanzee iPSCs from adult skin fibroblasts and reconstructed early neural development using in vitro differentiation culture conditions. Chimpanzee iPSCs were established using straightforward methods, namely, lipofection of plasmid vectors carrying human reprogramming factors, combined with maintenance in a comprehensive feeder-free culture. Ultimately, direct neurosphere formation culture induced rapid and efficient differentiation of neural stem cells from chimpanzee iPSCs. Time course analysis of neurosphere formation demonstrated ontogenetic changes in gene expression profiles and developmental potency along an early neural development path from epiblasts to radial glia. Our iPSC culture system is a potent tool for investigating the molecular and cellular foundation underlying chimpanzee early neural development and better understanding of human brain evolution.