Generation of mouse hippocampal brain organoids from primary embryonic neural stem cells

Summary: Here we present a protocol to generate standardized cerebral organoids with hippocampal regional specification using morphogen WNT3a. We describe steps for isolating mouse embryonic (E14.5) neural stem cells from the brain subgranular zone, preparing organoids samples for immunofluorescence...

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Main Authors: Francesca Ciarpella, Raluca Georgiana Zamfir, Alessandra Campanelli, Giulia Pedrotti, Marzia Di Chio, Emanuela Bottani, Ilaria Decimo
Format: Article
Language:English
Published: Elsevier 2023-09-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166723003805
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author Francesca Ciarpella
Raluca Georgiana Zamfir
Alessandra Campanelli
Giulia Pedrotti
Marzia Di Chio
Emanuela Bottani
Ilaria Decimo
author_facet Francesca Ciarpella
Raluca Georgiana Zamfir
Alessandra Campanelli
Giulia Pedrotti
Marzia Di Chio
Emanuela Bottani
Ilaria Decimo
author_sort Francesca Ciarpella
collection DOAJ
description Summary: Here we present a protocol to generate standardized cerebral organoids with hippocampal regional specification using morphogen WNT3a. We describe steps for isolating mouse embryonic (E14.5) neural stem cells from the brain subgranular zone, preparing organoids samples for immunofluorescence, calcium imaging, and metabolic profiling. This protocol can be used to generate mouse brain organoids for developmental studies, modeling disease, and drug screening. Organoids can be obtained in one month, thus providing a rapid tool for high-throughput data validation.For complete details on the use and execution of this protocol, please refer to Ciarpella et al. “Murine cerebral organoids develop network of functional neurons and hippocampal brain region identity”.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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spelling doaj.art-2eb4ba952a6f4341ae7fbbdf8259e9102023-07-16T04:19:13ZengElsevierSTAR Protocols2666-16672023-09-0143102413Generation of mouse hippocampal brain organoids from primary embryonic neural stem cellsFrancesca Ciarpella0Raluca Georgiana Zamfir1Alessandra Campanelli2Giulia Pedrotti3Marzia Di Chio4Emanuela Bottani5Ilaria Decimo6Section of Pharmacology, Department of Diagnostics and Public Health, University of Verona, Verona 37134, ItalySection of Pharmacology, Department of Diagnostics and Public Health, University of Verona, Verona 37134, ItalySection of Pharmacology, Department of Diagnostics and Public Health, University of Verona, Verona 37134, ItalySection of Pharmacology, Department of Diagnostics and Public Health, University of Verona, Verona 37134, ItalySection of Pharmacology, Department of Diagnostics and Public Health, University of Verona, Verona 37134, ItalySection of Pharmacology, Department of Diagnostics and Public Health, University of Verona, Verona 37134, ItalySection of Pharmacology, Department of Diagnostics and Public Health, University of Verona, Verona 37134, Italy; Corresponding authorSummary: Here we present a protocol to generate standardized cerebral organoids with hippocampal regional specification using morphogen WNT3a. We describe steps for isolating mouse embryonic (E14.5) neural stem cells from the brain subgranular zone, preparing organoids samples for immunofluorescence, calcium imaging, and metabolic profiling. This protocol can be used to generate mouse brain organoids for developmental studies, modeling disease, and drug screening. Organoids can be obtained in one month, thus providing a rapid tool for high-throughput data validation.For complete details on the use and execution of this protocol, please refer to Ciarpella et al. “Murine cerebral organoids develop network of functional neurons and hippocampal brain region identity”.1 : 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/S2666166723003805Cell BiologyModel OrganismsNeuroscienceStem CellsCell DifferentiationOrganoids
spellingShingle Francesca Ciarpella
Raluca Georgiana Zamfir
Alessandra Campanelli
Giulia Pedrotti
Marzia Di Chio
Emanuela Bottani
Ilaria Decimo
Generation of mouse hippocampal brain organoids from primary embryonic neural stem cells
STAR Protocols
Cell Biology
Model Organisms
Neuroscience
Stem Cells
Cell Differentiation
Organoids
title Generation of mouse hippocampal brain organoids from primary embryonic neural stem cells
title_full Generation of mouse hippocampal brain organoids from primary embryonic neural stem cells
title_fullStr Generation of mouse hippocampal brain organoids from primary embryonic neural stem cells
title_full_unstemmed Generation of mouse hippocampal brain organoids from primary embryonic neural stem cells
title_short Generation of mouse hippocampal brain organoids from primary embryonic neural stem cells
title_sort generation of mouse hippocampal brain organoids from primary embryonic neural stem cells
topic Cell Biology
Model Organisms
Neuroscience
Stem Cells
Cell Differentiation
Organoids
url http://www.sciencedirect.com/science/article/pii/S2666166723003805
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