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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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724007214 |
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| author | Dan Dou Erika L.F. Holzbaur C. Alexander Boecker |
| author_facet | Dan Dou Erika L.F. Holzbaur C. Alexander Boecker |
| author_sort | Dan Dou |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-abad15adb027452c8da12c075d060a1b |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-abad15adb027452c8da12c075d060a1b2025-01-13T04:19:11ZengElsevierSTAR Protocols2666-16672025-03-0161103556Protocol for live imaging of axonal transport in iPSC-derived iNeuronsDan Dou0Erika L.F. Holzbaur1C. Alexander Boecker2Department of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Neuroscience Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USADepartment of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Neuroscience Graduate Group, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USADepartment of Neurology, University Medical Center Goettingen, 37077 Goettingen, Germany; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166724007214cell biologymicroscopystem cells |
| spellingShingle | Dan Dou Erika L.F. Holzbaur C. Alexander Boecker Protocol for live imaging of axonal transport in iPSC-derived iNeurons STAR Protocols cell biology microscopy stem cells |
| title | Protocol for live imaging of axonal transport in iPSC-derived iNeurons |
| title_full | Protocol for live imaging of axonal transport in iPSC-derived iNeurons |
| title_fullStr | Protocol for live imaging of axonal transport in iPSC-derived iNeurons |
| title_full_unstemmed | Protocol for live imaging of axonal transport in iPSC-derived iNeurons |
| title_short | Protocol for live imaging of axonal transport in iPSC-derived iNeurons |
| title_sort | protocol for live imaging of axonal transport in ipsc derived ineurons |
| topic | cell biology microscopy stem cells |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724007214 |
| work_keys_str_mv | AT dandou protocolforliveimagingofaxonaltransportinipscderivedineurons AT erikalfholzbaur protocolforliveimagingofaxonaltransportinipscderivedineurons AT calexanderboecker protocolforliveimagingofaxonaltransportinipscderivedineurons |