Journal
DIFFERENTIATION
Volume 91, Issue 4-5, Pages 28-41Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.diff.2016.01.005
Keywords
Subependymal zone; Ventricular-subventricular zone; Neural stem cell; Neurogenic niches; Adult neurogenesis; Neurosphere assay; Stem cell culture
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Funding
- Ministerio de Economia y Competitividad (MINECO) [SAF2014-54581]
- CIBERNED [CB06/05/0086]
- RETIC Tercel [RD12/0019/0008]
- Generalitat Valenciana [PROMETEOII/2013/020, ISIC 2012/010]
- MINECO [SAF2012-40107]
- Fundacion BBVA
- Fundacion Botin-Banco de Santander
- MECDis FPU pre-doctoral fellowship
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Individual cells dissected from the subependymal neurogenic niche of the adult mouse brain proliferate in medium containing basic fibroblast growth factor (bFGF) and/or epidermal growth factor (EGF) as mitogens, to produce multipotent clonal aggregates called neurospheres. These cultures constitute a powerful tool for the study of neural stem cells (NSCs) provided that they allow the analysis of their features and potential capacity in a controlled environment that can be modulated and monitored more accurately than in vivo. Clonogenic and population analyses under mitogen addition or withdrawal allow the quantification of the self-renewing and multilineage potency of these cells and the identification of the mechanisms involved in these properties. Here, we describe a set of procedures developed and/or modified by our group including several experimental options that can be used either independently or in combination for the ex vivo assessment of cell properties of NSCs obtained from the adult subependymal niche. (C) 2016 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.
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