The disclosure reports on the identification and isolation of adult mouse lateral ventricle subventricular zone (SVZ) neurosphere initiating cells (NICs) by flow cytometry on the basis of GlastmidEGFRhighPlexinB2highCD24-/low04/PSA-NCAM-/lowTer119/CD45- markers (GEPCOT cells). These cells are highly mitotic and short-lived in vivo based on fate-mapping with Ascll CreERT2 and Dlϰl CreERT2 . In contrast, pre-GEPCOT cells were quiescent, expressed higher Glast, and lower EGFR and PlexinB2. Pre-GEPCOT cells could not form neurospheres but expressed the stem cell markers Glast-CreER T , GFAP-CreER T2 , Soϰ2 CreERT2 , and Glil CreERT2 and were long-lived in vivo. While GEPCOT NICs were ablated by temozolomide, pre-GEPCOT cells survived and repopulated the SVZ. Conditional deletion of the Bmi-1 polycomb protein depleted pre-GEPCOT and GEPCOT cells, though pre-GEPCOT cells were more dependent upon Bmi-1 for p16 Ink4a repression. These data distinguish quiescent NSCs from NICs and make it possible to study their properties in vivo.
IDENTIFICATION AND ISOLATION OF NEURAL STEM CELLS AND NEUROSPHERE INITIATING CELLS