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(a) principal: Ortega, Felipe
• Data de Publicação: 2011
• Outros Autores: Costa, Marcos Romualdo, Simon-Ebert, Tatiana, Schroeder, Timm, Götz, Magdalena
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UFRN
• Texto Completo: https://repositorio.ufrn.br/jspui/handle/123456789/23135
• topic: adult neural stem cells; single-cell level; cell culture; mouse

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.