Rizalar Filiz Sila, Haucke Volker
Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Berlin, Germany.
Department of Biology, Chemistry, Pharmacy, Freie Universität, Berlin, Germany.
Methods Mol Biol. 2025;2910:27-36. doi: 10.1007/978-1-0716-4446-1_2.
Generation of human induced pluripotent stem cells (iPSCs) provided a unique platform for human brain development studies, in vitro disease modeling, and therapeutic strategy development. Human stem cells can be rapidly and efficiently differentiated into several distinct subpopulations of brain cells. These stem cell-derived systems are gaining acceptance as a viable alternative in the neuroscience field as they can mimic interactions between various brain cells, and help recapitulate brain regions with specific functions. Here, we describe a method to generate functional, postmitotic, excitatory cortical-like neurons from iPSCs by expressing the NGN2 transgene from a stably integrated doxycycline-inducible promoter. These induced neurons (iNs) can be utilized to study the development and function of human cortical neurons. They also allow studying disease mechanisms by comparing normal and pathophysiological conditions and enable reliable screens for testing of therapeutic approaches.
人类诱导多能干细胞(iPSC)的产生为人类大脑发育研究、体外疾病建模和治疗策略开发提供了一个独特的平台。人类干细胞可以快速有效地分化为几个不同的脑细胞亚群。这些干细胞衍生系统作为神经科学领域一种可行的替代方法正逐渐被接受,因为它们可以模拟各种脑细胞之间的相互作用,并有助于重现具有特定功能的脑区。在这里,我们描述了一种通过从稳定整合的强力霉素诱导型启动子表达NGN2转基因,从iPSC生成功能性、有丝分裂后、兴奋性皮质样神经元的方法。这些诱导神经元(iN)可用于研究人类皮质神经元的发育和功能。它们还允许通过比较正常和病理生理条件来研究疾病机制,并能够进行可靠的筛选以测试治疗方法。