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2012
DOI: 10.1038/nprot.2012.123
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Isolation of multipotent neural stem or progenitor cells from both the dentate gyrus and subventricular zone of a single adult mouse

Abstract: In adult mammals, the subventricular zone of the lateral ventricles (SVZ) and the subgranular zone of the dentate gyrus (DG) demonstrate ongoing neurogenesis, and multipotent neural stem/progenitor cells (NSCs) in these two regions exhibit different intrinsic properties. However, investigation of the mechanisms underlying such differences has been limited by a lack of efficient methods for isolating NSCs, particularly from the adult DG. Here we describe a protocol that enables us to isolate self-renewing and m… Show more

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Cited by 155 publications
(138 citation statements)
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“…To obtain a single cell suspension, a mechanical dissociation was performed by passing the tissue fragments through a glass pipette. Single cells were seeded in a six-well plate in serum-free neurobasal medium (Miltenyi Biotech, Germany) supplemented with B27 and 2 mM glutamine (Gibco, Germany), 2 lg/ml heparin (Sigma, Germany), 20 ng/ml epidermal growth factor (R&D Systems, Germany) and 20 ng/ml basic fibroblast growth factor (CellSystems, Germany), 100 lg/ml penicillin, and 100 U/ ml streptomycin (Guo et al 2012). After 3 days of cultivation, the medium was changed.…”
Section: Methodsmentioning
confidence: 99%
“…To obtain a single cell suspension, a mechanical dissociation was performed by passing the tissue fragments through a glass pipette. Single cells were seeded in a six-well plate in serum-free neurobasal medium (Miltenyi Biotech, Germany) supplemented with B27 and 2 mM glutamine (Gibco, Germany), 2 lg/ml heparin (Sigma, Germany), 20 ng/ml epidermal growth factor (R&D Systems, Germany) and 20 ng/ml basic fibroblast growth factor (CellSystems, Germany), 100 lg/ml penicillin, and 100 U/ ml streptomycin (Guo et al 2012). After 3 days of cultivation, the medium was changed.…”
Section: Methodsmentioning
confidence: 99%
“…The primary aNPCs were prepared as previously described (Guo et al, 2012; Pan et al, 2013) from the SGZ of the DG from 6-7 week-old male C57BL/6J mice (Taconic, Hudson, NY). The solutions and media used during the aNPC isolation were filter sterilized.…”
Section: Methodsmentioning
confidence: 99%
“…Adult NSPCs can be isolated from neurogenic niches of adult CNSsubventricular zone (SVZ) or subgranular zone of hippocampal dentate gyrus [17]. In this case, the source availability is also the main limiting factor, which is further complicated by the difficulty to isolate and successfully maintain these cells in vitro [18][19][20].…”
Section: Sources Of Neural Stem Cellsmentioning
confidence: 99%