Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.
Stanley Center for Psychiatric Research, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
Cell Rep. 2018 May 22;23(8):2509-2523. doi: 10.1016/j.celrep.2018.04.066.
Transcription factor programming of pluripotent stem cells (PSCs) has emerged as an approach to generate human neurons for disease modeling. However, programming schemes produce a variety of cell types, and those neurons that are made often retain an immature phenotype, which limits their utility in modeling neuronal processes, including synaptic transmission. We report that combining NGN2 programming with SMAD and WNT inhibition generates human patterned induced neurons (hpiNs). Single-cell analyses showed that hpiN cultures contained cells along a developmental continuum, ranging from poorly differentiated neuronal progenitors to well-differentiated, excitatory glutamatergic neurons. The most differentiated neurons could be identified using a CAMK2A::GFP reporter gene and exhibited greater functionality, including NMDAR-mediated synaptic transmission. We conclude that utilizing single-cell and reporter gene approaches for selecting successfully programmed cells for study will greatly enhance the utility of hpiNs and other programmed neuronal populations in the modeling of nervous system disorders.
多能干细胞（PSCs）的转录因子编程已成为生成用于疾病建模的人类神经元的一种方法。然而，编程方案会产生多种细胞类型，而且生成的那些神经元往往保留不成熟的表型，这限制了它们在模拟神经元过程（包括突触传递）中的应用。我们报告称，将 NGN2 编程与 SMAD 和 WNT 抑制相结合可产生人类模式诱导神经元（hpiNs）。单细胞分析表明，hpiN 培养物中存在沿着发育连续体的细胞，范围从分化不良的神经元祖细胞到分化良好的兴奋性谷氨酸能神经元。最分化的神经元可以使用 CAMK2A::GFP 报告基因进行鉴定，并且表现出更高的功能，包括 NMDA 受体介导的突触传递。我们得出结论，利用单细胞和报告基因方法选择成功编程的细胞进行研究将极大地提高 hpiNs 和其他编程神经元群体在模拟神经系统疾病中的效用。
