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Título: Using an adherent cell culture of the mouse subependymal zone to study the behavior of adult neural stem cells on a single-cell level
Autor(es): Ortega, Felipe
Costa, Marcos R
Simon-Ebert, Tatiana
Schroeder, Timm
Götz, Magdalena
Palavras-chave: adult neural stem cells;single-cell level;cell culture;mouse
Data do documento: 3-Nov-2011
Resumo: A comprehensive understanding of the cell biology of adult neural stem cells (aNSCNSCNSCs) requires direct observation of aNSCNSCNSC division and lineage progression in the absence of niche-dependent signals. Here we describe a culture preparation of the adult mouse subependymal zone (SESEZ), which allows for continuous single-cell tracking of aNSCNSCNSC behavior. The protocol involves the isolation (~3 h) and culture of cells from the adult SESEZ at low density in the absence of mitogenic growth factors in chemically defined medium and subsequent live imaging using time-lapse video microscopy (5–7 d); these steps are followed by postimaging immunocytochemistry to identify progeny (~7 h). This protocol enables the observation of the progression from slow-dividing aNSCNSCNSCs of radial/astroglial identity up to the neuroblast stage, involving asymmetric and symmetric cell divisions of distinct fast-dividing precursors. This culture provides an experimental system for studying instructive or permissive effects of signal molecules on aNSCNSCNSC modes of cell division and lineage progression.
ISSN: 1754-2189
Aparece nas coleções:ICe - Artigos publicados em periódicos

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