诱导多能干细胞衍生神经元中线粒体功能的高内涵分析

High-Content Analysis of Mitochondrial Function in iPSC-Derived Neurons.

作者信息

Little Daniel, Luft Christin, Mosaku Olukunbi, Ketteler Robin, Devine Michael J, Gissen Paul

机构信息

MRC Laboratory for Molecular Cell Biology, University College London, London, UK.

Great Ormond Street Institute of Child Health, University College London, London, UK.

出版信息

Methods Mol Biol. 2019;1994:175-184. doi: 10.1007/978-1-4939-9477-9_16.

DOI:10.1007/978-1-4939-9477-9_16

PMID:31124115

Abstract

Mitochondrial dysfunction is linked to many neurological diseases; therefore, the ability to measure mitochondrial function is of great use for researching disease and testing potential therapeutics. Here we describe a high-content assay to simultaneously measure mitochondrial membrane potential, morphology and cell viability in iPSC-derived neurons. Neurons are seeded into plates suitable for fluorescent microscopy, stained with the mitochondrial membrane potential-dependent dye TMRM, cytoplasmic dye Calcein AM, and nuclear stain Hoechst 33342. Images are acquired in live cells and analyzed using automated image analysis software.

摘要

线粒体功能障碍与许多神经疾病相关；因此，测量线粒体功能的能力对于疾病研究和潜在治疗方法的测试非常有用。在此，我们描述了一种高内涵分析方法，用于同时测量诱导多能干细胞衍生神经元中的线粒体膜电位、形态和细胞活力。将神经元接种到适合荧光显微镜观察的培养板中，用依赖线粒体膜电位的染料四甲基罗丹明甲酯（TMRM）、细胞质染料钙黄绿素乙酰甲酯（Calcein AM）和核染料 Hoechst 33342 进行染色。在活细胞中采集图像，并使用自动图像分析软件进行分析。