Isolation of multipotent neural stem or progenitor cells from both the dentate gyrus and subventricular zone of a single adult mouse.
In adult mammals, the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone of the dentate gyrus (DG) show ongoing neurogenesis, and multipotent neural stem or 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 2-3 h. As 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.
Waisman Center, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Weixiang
W
Guo
Natalie E
NE
Patzlaff
Emily M
EM
Jobe
Xinyu
X
Zhao
D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D018891:Dentate Gyrus / Q000166:cytology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D020547:Lateral Ventricles / Q000166:cytology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D042282:Microdissection / Q000379:methods*; D039902:Multipotent Stem Cells / Q000166:cytology*; D058953:Neural Stem Cells / Q000166:cytology*
D049452:Green Fluorescent Proteins