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Open Access Green as soon as Postprint is submitted to ZB.
Using an adherent cell culture of the mouse subependymal zone to study the behavior of adult neural stem cells on a single-cell level.
Nat. Protoc. 6, 1847-1859 (2011)
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 (∼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 (∼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.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
no keywords
ISSN (print) / ISBN
1754-2189
e-ISSN
1750-2799
Journal
Nature Protocols
Quellenangaben
Volume: 6,
Issue: 12,
Pages: 1847-1859
Publisher
Nature Publishing Group
Non-patent literature
Publications
Reviewing status
Peer reviewed