Isolation of multipotent neural stem or progenitor cells from both the dentate gyrus and subventricular zone of a single adult mouse.
Isolation of multipotent neural stem or progenitor cells from both the dentate gyrus and subventricular zone of a single adult mouse.
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DOI：
10.1038/nprot.2012.123
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发表时间：
2012-11
期刊：
影响因子：
14.8
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中科院分区：
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在成年哺乳动物中，侧脑室室下区(SVZ)和齿状回颗粒下区(DG)显示出持续的神经发生，这两个区域的多潜能神经干/祖细胞(NSCs)显示出不同的内在特性。然而，由于缺乏有效的方法来分离神经干细胞，尤其是从成人DG中分离神经干细胞，对这种差异的机制的研究一直受到限制。在这里，我们描述了一种方案，使我们能够从同一成年小鼠的SVZ和DG中分离出自我更新和多潜能的神经干细胞。该方案包括从成年小鼠脑中显微解剖SVZ和DG，从特定区域分离NSCs，并在体外培养NSCs。整个过程需要2到3个小时。由于每个细胞分离程序只需要一只小鼠，因此该方案对于小鼠数量有限的研究特别有用，例如包含多个基因修饰的小鼠。
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 multipotent NSCs from the SVZ and the DG of the same adult mouse. The protocol involves the microdissection of the SVZ and DG from one adult mouse brain, isolation of NSCs from specific regions, and cultivation of NSCs in vitro. The entire procedure takes 2 to 3 hours. Since only one mouse is needed for each cell isolation procedure, this protocol will be particularly useful for studies with limited availability of mice, such as mice that contain multiple genetic modifications.