Flow cytometric analysis of neural stem cells in the developing and adult mouse brain

Ayako Murayama, Yumi Matsuzaki, Ayano Kawaguchi, Takuya Shimazaki, Hideyuki Okano

Research output: Contribution to journalArticle

106 Citations (Scopus)

Abstract

Despite recent progress in the neural stem cell biology, their cellular characteristics have not been described well. We investigated various characteristics of neural stem cells (NSCs) in vivo during CNS development, using FACS to identify the NSCs. We first examined stage-dependent changes in the physical parameters, using forward scatter (FSC) and side scatter (SSC) profiles, of NSCs from the developing striatum, where they appear to be active throughout the life of mammals. NSCs were divided into several fractions according to their FSC/SSC profile. With development, their number decreased in the FSChigh fractions but increased in the FSClow/SSChigh fraction, whereas NSCs were significantly concentrated in the fraction containing the largest cells (about 20 μm in diameter) at any stage, which were mostly the cells with the highest nestin-enhancer activity. Furthermore, we demonstrated that, at all stages examined, the "side population" (SP), defined as the Hoechst 33342 low/negative fraction, which is known to be a stem cell-enriched population in bone marrow, was also enriched for Notch1-positive immature neural cells (about 60%) from the developing striatum. However, these immature SP cells were not detected in the large-cell fraction, however, but were concentrated instead in the FSClow/mid fractions. FACS analysis showed that SP cells from adults were included to some extent in the CD24low/ PNAlow fraction, where NSCs were greatly concentrated. Collectively, the characteristics of NSCs were not uniform and changed developmentally.

Original languageEnglish
Pages (from-to)837-847
Number of pages11
JournalJournal of neuroscience research
Volume69
Issue number6
DOIs
Publication statusPublished - 2002 Sep 15

    Fingerprint

Keywords

  • CD24
  • Fluorescent activated cell sorting
  • Forward scatter
  • Nestin-EGFP transgenic mice
  • Neural stem cells
  • Neurosphere
  • Notch1
  • Peanut agglutinin
  • Side population
  • Side scatter

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this