Nestin-EGFP transgenic mice: Visualization of the self-renewal and multipotency of CNS stem cells

Ayano Kawaguchi, Takaki Miyata, Kazunobu Sawamoto, Noriko Takashita, Ayako Murayama, Wado Akamatsu, Masaharu Ogawa, Masaru Okabe, Yasuo Tano, Steven A. Goldman, Hideyuki Okano

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259 Citations (Scopus)

Abstract

We generated transgenic mice carrying enhanced green fluorescent protein (EGFP) under the control of the nestin second-intronic enhancer (E/nestin:EGFP). Flow cytometry followed by in vitro assays revealed that in situ EGFP expression in the embryonic brain correlated with the mitotic index, the cogeneration of both neurons and gila, and the frequency of neurosphere formation in vitro. High-level EGFP expressors derived from embryos included a distinct subpopulation of cells that were self-renewable and multipotent, criteria that define neural stern cells (NSCs). Such cells were largely absent among lower-level or non-EGFP expressors, thereby permitting us to enrich for NSCs using EGFP expression level. In adults, although E/nestin:EGFP-positive cells included the NSC population, the frequency of neurosphere formation did not correlate directly with the level of EGFP expression. However, moderately EGFP-expressing cells in adults gained EGFP intensity when they formed neurospheres, suggesting embryonic and adult NSCs exist in different microenvironments in vivo.

Original languageEnglish
Pages (from-to)259-273
Number of pages15
JournalMolecular and Cellular Neuroscience
Volume17
Issue number2
DOIs
Publication statusPublished - 2001 Jan 1
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience
  • Cell Biology

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    Kawaguchi, A., Miyata, T., Sawamoto, K., Takashita, N., Murayama, A., Akamatsu, W., Ogawa, M., Okabe, M., Tano, Y., Goldman, S. A., & Okano, H. (2001). Nestin-EGFP transgenic mice: Visualization of the self-renewal and multipotency of CNS stem cells. Molecular and Cellular Neuroscience, 17(2), 259-273. https://doi.org/10.1006/mcne.2000.0925