Prospective identification of functionally distinct stem cells and neurosphere-initiating cells in adult mouse forebrain

Elife. 2014 May 7;3:e02669. doi: 10.7554/eLife.02669.


Neurosphere formation is commonly used as a surrogate for neural stem cell (NSC) function but the relationship between neurosphere-initiating cells (NICs) and NSCs remains unclear. We prospectively identified, and isolated by flow cytometry, adult mouse lateral ventricle subventricular zone (SVZ) NICs as Glast(mid)EGFR(high)PlexinB2(high)CD24(-/low)O4/PSA-NCAM(-/low)Ter119/CD45(-) (GEPCOT) cells. They were 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-GEPCOT cells could not form neurospheres but expressed the stem cell markers Slc1a3-CreER(T), GFAP-CreER(T2), Sox2(CreERT2), and Gli1(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 Cdkn2a (p16(Ink4a)) repression. Our data distinguish quiescent NSCs from NICs and make it possible to study their properties in vivo.DOI:

Keywords: Bmi-1; fate-mapping; forebrain; prospective identification; stem cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aging / metabolism*
  • Animals
  • Antimitotic Agents / pharmacology
  • Cell Proliferation
  • Cell Separation
  • Dacarbazine / analogs & derivatives
  • Dacarbazine / pharmacology
  • Glial Fibrillary Acidic Protein / metabolism
  • Integrases / metabolism
  • Mice, Inbred C57BL
  • Neural Stem Cells / cytology*
  • Neurogenesis / drug effects
  • Neuroglia / cytology
  • Neuroglia / metabolism
  • Phenotype
  • Polycomb Repressive Complex 1 / metabolism
  • Prosencephalon / cytology*
  • Proto-Oncogene Proteins / metabolism
  • Spheroids, Cellular / cytology*
  • Temozolomide


  • Antimitotic Agents
  • Bmi1 protein, mouse
  • Glial Fibrillary Acidic Protein
  • Proto-Oncogene Proteins
  • Dacarbazine
  • Polycomb Repressive Complex 1
  • Cre recombinase
  • Integrases
  • Temozolomide