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

Annika Zink; Undine Haferkamp; Annika Wittich; Mathias Beller; Ole Pless; Alessandro Prigione

doi:10.1016/j.xpro.2022.101602

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

STAR Protoc. 2022 Aug 5;3(3):101602. doi: 10.1016/j.xpro.2022.101602. eCollection 2022 Sep 16.

Authors

Annika Zink¹, Undine Haferkamp², Annika Wittich², Mathias Beller³, Ole Pless², Alessandro Prigione⁴

Affiliations

¹ Department of General Pediatrics, Neonatology and Pediatric Cardiology, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany.
² Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, ScreeningPort, 22525 Hamburg, Germany.
³ Institute for Mathematical Modeling of Biological Systems, Heinrich Heine University, 40225 Düsseldorf, Germany.
⁴ Department of General Pediatrics, Neonatology and Pediatric Cardiology, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany. Electronic address: alessandro.prigione@hhu.de.

Abstract

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).

Keywords: Cell Biology; Cell-based Assays; Metabolism; Microscopy; Neuroscience; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

High-Throughput Screening Assays / methods
Humans
Induced Pluripotent Stem Cells* / metabolism
Mitochondria / physiology
Neurons
Pluripotent Stem Cells* / metabolism