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.

我们最亲近的近亲黑猩猩（Pan troglodytes）的进化发育生物学对于理解人类特征的起源至关重要。但是，由于技术和伦理上的限制，很难在体内获得黑猩猩的发育事件。诱导性多能干细胞（iPSC）提供了另一种体外模型系统，可通过克服体内研究的局限性来研究发育事件。在这里，我们从成年皮肤成纤维细胞中生成了黑猩猩iPSC，并使用体外重建了早期的神经发育分化培养条件。黑猩猩iPSC是使用直接方法建立的，即携带人重编程因子的质粒载体脂转染，并在无饲养层的全面培养中进行维护。最终，直接的神经球形成培养物诱导了黑猩猩iPSC的神经干细胞的快速有效分化。神经球形成的时程分析表明，沿从表皮细胞到radial神经胶质细胞的早期神经发育路径，基因表达谱和发育潜能的遗传学变化。我们的iPSC培养系统是研究黑猩猩早期神经发育和更好地了解人脑进化的分子和细胞基础的有效工具。