期刊
NATURE PROTOCOLS
卷 6, 期 12, 页码 1847-1859出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2011.404
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资金
- Deutsche Forschungsgemeinschaft [BE 4182/2-2, GO 640/9-2, SCHR 1142/1-2]
- Bundesministerium fur Bildung und Forschung
- Bavarian State Ministry of Sciences, Research and the Arts
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 (similar to 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 (similar to 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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