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Mice Post-natal Subventricular Zone Neurospheres: Derivation, Culture, Differentiation and Applications

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Working with Stem Cells

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Abstract

Neurogenesis is a highly regulated process, by which the brain is able to produce new neural cells mainly under normal conditions or after injury. Neurogenesis is restricted to specific areas in the brain, known as niches, where neural stem cells (NSCs) are generated. Two main regions have been described as neurogenic in mammalian brain, the subventricular zone (SVZ) in the anterolateral ventricles, and the subgranular zone (SGZ) of the dentate gyrus in the hippocampus. Under appropriate microenviromental circumstances, NSCs divide and give rise to neural progenitor cells (NPCs) that will generate mature neurons or glia. These precursors can be obtained from rodent animal models to produce neurospheres as an in vitro model for neurogenesis. Neurospheres are aggregates of NPC that grow in suspension and are capable of differentiating into every lineage of neural cells, including neurons and glia. This capacity shows the high potential of neurospheres as an in vitro model to study neurogenesis and alternatives methodologies to investigate neurodegenerative disorders. Here, we present an easy and highly reproducible protocol for working with SVZ neurospheres from post-natal mice.

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Acknowledgements

HU acknowledges grant support by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP Project No. 2012/50880-4) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq Project No. 486294/2012-9 and 467465/2014-2), Brazil. LSA’s doctoral thesis research is supported by a fellowship from FAPESP (Project No. 2013/25338-4). CS is recipient of a CAPES-FAPERJ Postdoc fellowship. SX postdoctoral research is supported by a fellowship from FCT-Portugal (SFRH/BPD/ 76642/2011). We would like to acknowledge Ana Moreira and Rui Rodrigues for their kind help with photos and images for Fig. 5.2 .

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Authors and Affiliations

  1. Department of Biochemistry, Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes 748, São Paulo, SP, 05508-000, Brazil

    Laura Sardá-Arroyo & Henning Ulrich

  2. Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, 21941-902, Brazil

    Clarissa Schitine

  3. Instituto de Farmacologia e Neurociências and Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Av. Prof. Egas Moniz, Lisbon, 1649-028, Portugal

    Sara Alves Xapelli

Authors
  1. Laura Sardá-Arroyo
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  2. Clarissa Schitine
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  3. Sara Alves Xapelli
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  4. Henning Ulrich
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Correspondence to Henning Ulrich .

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Sardá-Arroyo, L., Schitine, C., Xapelli, S.A., Ulrich, H. (2016). Mice Post-natal Subventricular Zone Neurospheres: Derivation, Culture, Differentiation and Applications. In: Working with Stem Cells. Springer, Cham. https://doi.org/10.1007/978-3-319-30582-0_5

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