Generation of human-induced pluripotent-stem-cell-derived cortical neurons for high-throughput imaging of neurite morphology and neuron maturation

Gautam Wali; Yan Li; Dad Abu-Bonsrah; Deniz Kirik; Clare L. Parish; Carolyn M. Sue

STAR Protocols (Jun 2023)

Generation of human-induced pluripotent-stem-cell-derived cortical neurons for high-throughput imaging of neurite morphology and neuron maturation

Gautam Wali,
Yan Li,
Dad Abu-Bonsrah,
Deniz Kirik,
Clare L. Parish,
Carolyn M. Sue

Affiliations

Gautam Wali: 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; Corresponding author
Yan Li: 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
Dad Abu-Bonsrah: The Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Parkville, VIC 3052, Australia; Department of Paediatrics, The University of Melbourne, Parkville, VIC 3052, Australia; Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Deniz Kirik: BRAINS Unit, BMC D11, Lund University, 22184 Lund, Sweden; Honorary Professorship at School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2006, Australia; Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Clare L. Parish: The Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Parkville, VIC 3052, Australia; Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Carolyn M. Sue: 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

Journal volume & issue: Vol. 4, no. 2
p. 102325

Abstract

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Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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