title: Isolation, culture and analysis of adult subependymal neural stem cells.
abstract: 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.
authors.affiliation: Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Spain; Departamento de Biología Celular and ERI BiotecMed, Universidad de Valencia, 46100, Spain.
authors.forename: Germán
authors.initials: G
authors.lastname: Belenguer
authors.affiliation: Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Spain; Departamento de Biología Celular and ERI BiotecMed, Universidad de Valencia, 46100, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Domingo-Muelas
authors.affiliation: Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Spain; Departamento de Biología Celular and ERI BiotecMed, Universidad de Valencia, 46100, Spain.
authors.forename: Sacri R
authors.initials: SR
authors.lastname: Ferrón
authors.affiliation: Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Spain; Departamento de Biología Celular and ERI BiotecMed, Universidad de Valencia, 46100, Spain. Electronic address: jm.morante@uv.es.
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Morante-Redolat
authors.affiliation: Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Spain; Departamento de Biología Celular and ERI BiotecMed, Universidad de Valencia, 46100, Spain. Electronic address: isabel.farinas@uv.es.
authors.forename: Isabel
authors.initials: I
authors.lastname: Fariñas
mesh_terms: D053687:Adult Stem Cells; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D004805:Ependyma / Q000166:cytology / Q000254:growth & development*; D006801:Humans; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology*; D055495:Neurogenesis / Q000235:genetics*; D009474:Neurons
keywords: Adult neurogenesis; Neural stem cell; Neurogenic niches; Neurosphere assay; Stem cell culture; Subependymal zone; Ventricular–subventricular zone