High-content screening of mitochondrial polarization in neural cells derived from human pluripotent stem cells

• Published in: STAR protocols Vol. 3; no. 3; p. 101602
• Main Authors: Zink, Annika, Haferkamp, Undine, Wittich, Annika, Beller, Mathias, Pless, Ole, Prigione, Alessandro
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.09.2022; Elsevier
• Subjects: Cell Biology; Cell-based Assays; High-Throughput Screening Assays - methods; Humans; Induced Pluripotent Stem Cells - metabolism; Metabolism; Microscopy; Mitochondria - physiology; Neurons; Neuroscience; Pluripotent Stem Cells - metabolism; Protocol; Stem Cells; Stem Cells; Neuroscience; Microscopy; Metabolism; Cell-based Assays; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101602

We present a high-content screening (HCS) protocol for quantifying mitochondrial activity in live neural cells from human induced pluripotent stem cells (iPSCs). The assessment is based on mitochondrial membrane potential, which is influenced by the efficiency of mitochondrial bioenergetics. We describe how to perform the analysis using both an HCS platform and the open-source software CellProfiler. The protocol can identify the mitochondrial fitness of human neurons and may be used to carry out high-throughput compound screenings in patient-derived neural cells. For complete details on the use and execution of this protocol, please refer to Lorenz et al. (2017) and Zink et al. (2020). [Display omitted] •High-content screening (HCS) of live neural cells derived from human iPSCs•Assessment of mitochondrial activity based on mitochondrial membrane potential (MMP)•Quantification using automated microscopic image analysis•Data analysis using both an HCS system and the open-source software CellProfiler Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. We present a high-content screening (HCS) protocol for quantifying mitochondrial activity in live neural cells from human induced pluripotent stem cells (iPSCs). The assessment is based on mitochondrial membrane potential, which is influenced by the efficiency of mitochondrial bioenergetics. We describe how to perform the analysis using both an HCS platform and the open-source software CellProfiler. The protocol can identify the mitochondrial fitness of human neurons and may be used to carry out high-throughput compound screenings in patient-derived neural cells.