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Isolation and Expansion of Neurospheres from Postnatal (P1−3) Mouse Neurogenic Niches
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Isolation and Expansion of Neurospheres from Postnatal (P1−3) Mouse Neurogenic Niches

Isolation and Expansion of Neurospheres from Postnatal (P1−3) Mouse Neurogenic Niches

DOI:
10.3791/60822-v

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10:05 min

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May 23, 2020

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, , , , , , ,
1Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina,Universidade de Lisboa, 2Instituto de Farmacologia e Neurociências, Faculdade de Medicina,Universidade de Lisboa, 3Instituto de Biologia Molecular, Faculdade de Medicina,Universidade de Lisboa

Chapters

  • 00:04Introduction
  • 00:59Postnatal Day 1-3 (P1-3) Mouse Brain Harvest
  • 02:28Subventricular Zone (SVZ)/Dentate Gyrus (DG) Microdissections
  • 04:22Tissue Dissociation
  • 05:27Postnatal Neural Stem Cell Expansion
  • 06:09Neurosphere Passaging
  • 07:10Results: Representative Neurosphere Expansion and Characterization
  • 09:21Conclusion

Summary

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In this article, we describe, in detail, a protocol for the generation of neurosphere cultures from postnatal mouse neural stem cells derived from the main mouse neurogenic niches. Neurospheres are used to identify neural stem cells from brain tissue allowing the estimation of precursor cell numbers. Moreover, these 3D structures can be plated in differentiative conditions, giving rise to neurons, oligodendrocytes and astrocytes, allowing the study of cell fate.

Tags

Keywords: Neurosphere AssayNeural Stem/progenitor CellsAdult NeurogenesisBrain DisordersAlzheimer’sParkinson’sMultiple SclerosisPostnatal Mouse BrainSubventricular ZoneDentate GyrusMicrodissectionHBSSMeningesCerebellumOlfactory Bulbs

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