Protocol for live imaging of axonal transport in iPSC-derived iNeurons

Summary: Studies of human induced pluripotent stem cell (iPSC)-derived neurons promise important insights into neurodegenerative diseases. Here, we present a protocol for live imaging of axonal transport in glutamatergic iPSC-derived neurons (iNeurons). We describe steps for the differentiation of i...

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Bibliographic Details
Main Authors: Dan Dou, Erika L.F. Holzbaur, C. Alexander Boecker
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724007214
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Summary:Summary: Studies of human induced pluripotent stem cell (iPSC)-derived neurons promise important insights into neurodegenerative diseases. Here, we present a protocol for live imaging of axonal transport in glutamatergic iPSC-derived neurons (iNeurons). We describe steps for the differentiation of iPSCs into iNeurons via PiggyBac-mediated neurogenin 2 (NGN2) delivery, iNeuron culture and transfection, and the acquisition and analysis of time-lapse images. Our protocol is optimized for the widely available catalog of KOLF2.1J iPSCs with mutations relevant to neurodegenerative diseases but is also applicable to other iPSC lines.For complete details on the use and execution of this protocol, please refer to Dou et al.1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667