用于神经突形态和神经元成熟高通量成像的人诱导多能干细胞衍生皮质神经元的生成
Generation of human-induced pluripotent-stem-cell-derived cortical neurons for high-throughput imaging of neurite morphology and neuron maturation.
作者信息
Wali Gautam, Li Yan, Abu-Bonsrah Dad, Kirik Deniz, Parish Clare L, Sue Carolyn M
机构信息
Neuroscience Research Australia and University of New South Wales, Sydney, NSW 2031, Australia; Kolling Institute for Medical Research and Department of Medicine, University of Sydney, Sydney, NSW 2065, Australia; Northern Clinical School, Sydney Medical School, University of Sydney, Sydney, NSW 2065, Australia; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA.
Neuroscience Research Australia and University of New South Wales, Sydney, NSW 2031, Australia; Kolling Institute for Medical Research and Department of Medicine, University of Sydney, Sydney, NSW 2065, Australia; Northern Clinical School, Sydney Medical School, University of Sydney, Sydney, NSW 2065, Australia.
出版信息
STAR Protoc. 2023 Jun 9;4(2):102325. doi: 10.1016/j.xpro.2023.102325.
High-throughput imaging allows in vitro assessment of neuron morphology for screening populations under developmental, homeostatic, and/or disease conditions. Here, we present a protocol to differentiate cryopreserved human cortical neuronal progenitors into mature cortical neurons for high-throughput imaging analysis. We describe the use of a notch signaling inhibitor to generate homogeneous neuronal populations at densities amenable to individual neurite identification. We detail neurite morphology assessment via measuring multiple parameters including neurite length, branches, roots, segments and extremities, and neuron maturation.
高通量成像能够在体外评估神经元形态，用于筛选处于发育、稳态和/或疾病状态下的细胞群体。在此，我们展示了一种将冷冻保存的人类皮质神经元祖细胞分化为成熟皮质神经元以进行高通量成像分析的方案。我们描述了使用一种Notch信号抑制剂来生成密度适合单个神经突识别的均匀神经元群体。我们详细介绍了通过测量多个参数（包括神经突长度、分支、根部、节段和末梢）以及神经元成熟度来评估神经突形态的方法。
Zotero 插件