High-content analysis of neuronal morphology in human iPSC-derived neurons

Selene Lickfett; Carmen Menacho; Annika Zink; Narasimha Swamy Telugu; Mathias Beller; Sebastian Diecke; Sidney Cambridge; Alessandro Prigione

doi:10.1016/j.xpro.2022.101567

High-content analysis of neuronal morphology in human iPSC-derived neurons

STAR Protoc. 2022 Aug 6;3(3):101567. doi: 10.1016/j.xpro.2022.101567. eCollection 2022 Sep 16.

Authors

Selene Lickfett¹, Carmen Menacho², Annika Zink², Narasimha Swamy Telugu³, Mathias Beller⁴, Sebastian Diecke³, Sidney Cambridge⁵, Alessandro Prigione⁶

Affiliations

¹ Department of General Pediatrics, Neonatology and Pediatric Cardiology, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany; Department of Anatomy II, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany.
² Department of General Pediatrics, Neonatology and Pediatric Cardiology, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany.
³ Max Delbrück Center for Molecular Medicine (MDC) and Berlin Institute of Health (BIH), 13125 Berlin, Germany.
⁴ Institute for Mathematical Modeling of Biological Systems, Heinrich Heine University, 40225 Düsseldorf, Germany.
⁵ Department of Anatomy II, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany; Dr. Senckenbergische Anatomie, Goethe-Universität Frankfurt, 60590 Frankfurt am Main, Germany.
⁶ 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 analysis (HCA) protocol for monitoring the outgrowth capacity of human neurons derived from induced pluripotent stem cells (iPSCs). We describe steps to perform HCA imaging, followed by quantifying the morphology of dendrites and axons within a high-throughput system to evaluate neurons obtained through various differentiation approaches. This protocol can be used to screen for modulators of neuronal morphogenesis or neurotoxicity. The approach can be applied to patient-derived iPSCs to identify patient-specific defects and possible therapeutic strategies. For complete details on the use and execution of this protocol, please refer to Zink et al. (2020) and Inak et al. (2021). The protocol can be used in combination with Zink et al. (2022).

Keywords: Cell Differentiation; High Throughput Screening; Microscopy; Neuroscience; Stem Cells.

Publication types

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

MeSH terms

Cell Differentiation / physiology
Humans
Induced Pluripotent Stem Cells*
Neurons
Neurotoxicity Syndromes*