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

Journal of Visualized Experiments : JoVE
Rita SoaresSara Xapelli

Abstract

The neurosphere assay is an extremely useful in vitro technique for studying the inherent properties of neural stem/progenitor cells (NSPCs) including proliferation, self-renewal and multipotency. In the postnatal and adult brain, NSPCs are mainly present in two neurogenic niches: the subventricular zone (SVZ) lining the lateral ventricles and the subgranular zone of the hippocampal dentate gyrus (DG). The isolation of the neurogenic niches from postnatal brain allows obtaining a higher amount of NSPCs in culture with a consequent advantage of higher yields. The close contact between cells within each neurosphere creates a microenvironment that may resemble neurogenic niches. Here, we describe, in detail, how to generate SVZ- and DG-derived neurosphere cultures from 1-3-day-old (P1-3) mice, as well as passaging, for neurosphere expansion. This is an advantageous approach since the neurosphere assay allows a fast generation of NSPC clones (6-12 days) and contributes to a significant reduction in the number of animal usage. By plating neurospheres in differentiative conditions, we can obtain a pseudomonolayer of cells composed of NSPCs and differentiated cells of different neural lineages (neurons, astrocytes and oligodendrocytes...Continue Reading

Citations

May 2, 2021·Journal of Neuroscience Methods·James A CarrollCathryn L Haigh

❮ Previous
Next ❯

Related Concepts

Related Feeds

Astrocytes

Astrocytes are glial cells that support the blood-brain barrier, facilitate neurotransmission, provide nutrients to neurons, and help repair damaged nervous tissues. Here is the latest research.

Related Papers

International Journal of Developmental Neuroscience : the Official Journal of the International Society for Developmental Neuroscience
Tao SunHua Xin
International Journal of Developmental Neuroscience : the Official Journal of the International Society for Developmental Neuroscience
Ema F TorradoAna S Falcão
© 2022 Meta ULC. All rights reserved