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Ortega, F.* ; Costa, M.R.* ; Simon-Ebert, T.* ; Schroeder, T. ; Götz, M. ; Berninger, B.

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)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
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
Language english
Publication Year 2011
HGF-reported in Year 2011
ISSN (print) / ISBN 1754-2189
e-ISSN 1750-2799
Journal Nature Protocols
Quellenangaben Volume: 6, Issue: 12, Pages: 1847-1859 Article Number: , Supplement: ,
Publisher Nature Publishing Group
Reviewing status Peer reviewed
Institute(s) Research Unit Stem Cell Dynamics (SCD)
Institute of Stem Cell Research (ISF)
POF-Topic(s) 30504 - Mechanisms of Genetic and Environmental Influences on Health and Disease
30204 - Cell Programming and Repair
Research field(s) Stem Cell and Neuroscience
PSP Element(s) G-501200-001
G-500800-001
PubMed ID 22051798
DOI 10.1038/nprot.2011.404
Scopus ID 80955152015
Erfassungsdatum 2011-11-25
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