Dimitrion Peter, Zhi Yun, Clayton Dennis, Apodaca Gerard L, Wilcox Madeleine R, Johnson Jon W, Nimgaonkar Vishwajit, D'Aiuto Leonardo
Department of Psychiatry, Western Psychiatric Institute and Clinic, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
Department of Pharmacology and Pharmaceutical Sciences, School of Medicine, Tsinghua University, Beijing, China.
Mol Neuropsychiatry. 2017 Jul;3(1):28-36. doi: 10.1159/000476034. Epub 2017 Jun 17.
Induced pluripotent stem cell (iPSC)-based technologies offer an unprecedented possibility to investigate defects occurring during neuronal differentiation in neuropsychiatric and neurodevelopmental disorders, but the density and intricacy of intercellular connections in neuronal cultures challenge currently available analytic methods. Low-density neuronal cultures facilitate the morphometric and functional analysis of neurons. We describe a differentiation protocol to generate low-density neuronal cultures (∼2,500 neurons/cm) from human iPSC-derived neural stem cells/early neural progenitor cells. We generated low-density cultures using cells from 3 individuals. We also evaluated the morphometric features of neurons derived from 2 of these individuals, one harboring a microdeletion on chromosome 15q11.2 and the other without the microdeletion. An approximately 7.5-fold increase in the density of dendritic filopodia was observed in the neurons with the microdeletion, consistent with previous reports. Low-density neuronal cultures enable facile and unbiased comparisons of iPSC-derived neurons from different individuals or clones.
基于诱导多能干细胞（iPSC）的技术为研究神经精神疾病和神经发育障碍中神经元分化过程中出现的缺陷提供了前所未有的可能性，但神经元培养物中细胞间连接的密度和复杂性对目前可用的分析方法提出了挑战。低密度神经元培养有助于对神经元进行形态计量和功能分析。我们描述了一种从人iPSC衍生的神经干细胞/早期神经祖细胞生成低密度神经元培养物（约2500个神经元/cm²）的分化方案。我们使用来自3个个体的细胞生成了低密度培养物。我们还评估了其中2个个体来源的神经元的形态计量特征，一个在染色体15q11.2上有微缺失，另一个没有微缺失。与之前的报道一致，在有微缺失的神经元中观察到树突丝状伪足密度增加了约7.5倍。低密度神经元培养能够轻松且无偏地比较来自不同个体或克隆的iPSC衍生神经元。
