title: Using an adherent cell culture of the mouse subependymal zone to study the behavior of adult neural stem cells on a single-cell level.
abstract: A comprehensive understanding of the cell biology of adult neural stem cells (aNSCs) requires direct observation of aNSC division and lineage progression in the absence of niche-dependent signals. Here we describe a culture preparation of the adult mouse subependymal zone (SEZ), which allows for continuous single-cell tracking of aNSC behavior. The protocol involves the isolation (approximately 3 h) and culture of cells from the adult SEZ 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 (approximately 7 h). This protocol enables the observation of the progression from slow-dividing aNSCs 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 aNSC modes of cell division and lineage progression.
authors: Department of Physiological Genomics, Institute of Physiology, Ludwig-Maximilians University Munich, Germany.
authors: Felipe
authors: F
authors: Ortega
authors: Marcos R
authors: MR
authors: Costa
authors: Tatiana
authors: T
authors: Simon-Ebert
authors: Timm
authors: T
authors: Schroeder
authors: Magdalena
authors: M
authors: Götz
authors: Benedikt
authors: B
authors: Berninger
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002455:Cell Division; D019070:Cell Lineage; D017930:Genes, Reporter; D051379:Mice; D008810:Mice, Inbred C57BL; D058953:Neural Stem Cells / Q000166:cytology*; D012984:Software; D059008:Time-Lapse Imaging; D014741:Video Recording