IDENTIFICATION AND ISOLATION OF NEURAL STEM CELLS AND NEUROSPHERE INITIATING CELLS

摘要

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 Glast^midEGFR^highPlexinB2^highCD24^−/lowO4/PSA-NCAM^−/lowTer-119/CD45⁻ markers (GEPCOT cells). These cells are highly mitotic and short-lived in vivo based on fate-mapping with Ascl1^CreERT2 and Dlx1^CreERT2. In contrast, pre-GEPCOT cells were quiescent, expressed higher Glast, and lower EGFR and PlexinB2. Pre-GEP-COT cells could not form neurospheres but expressed the stem cell markers Glast-CreER^T, GFAP-CreER^T2, Sox2^CreERT2, and Gli1^C-reERT2 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.