High-yield selection and extraction of two promoter-defined phenotypes of neural stem cells from the fetal human brain

H. Michael Keyoung, Neeta S. Roy, Abdellatif Benraiss, Abner Louissaint, Akira Suzuki, Mitsuhiro Hashimoto, William K. Rashbaum, Hideyuki Okano, Steven A. Goldman

Research output: Contribution to journalArticle

141 Citations (Scopus)

Abstract

Neural stem and precursor cells reside in the ventricular lining of the fetal forebrain, and may provide a cellular substrate for brain repair. To selectively identify and extract these cells, we infected dissociated fetal human brain cells with adenoviruses bearing the gene for green fluorescence protein (GFP), placed under the control of enhancer/promoters for two genes (nestin and musashil) that are expressed in uncommitted neuroepithelial cells. The cells were then sorted by fluorescence-activated cell sorting (FACS) on the basis of E/nestin- or P/musashi1-driven GFP expression. Both P/musashi1:hGFP- and E/nestin:EGFP-sorted cells were multipotent: limiting dilution with clonal expansion as neurospheres, in tandem with retroviral lineage analysis and xenograft to E17 and P0-2 rat forebrain, revealed that each phenotype was able to both self-renew and co-generate neurons and glia. Thus, fluorescent genes placed under the control of early neural promoters allow neural stem cells to be specifically targeted, isolated, and substantially enriched from the fetal human brain.

Original languageEnglish
Pages (from-to)843-850
Number of pages8
JournalNature Biotechnology
Volume19
Issue number9
DOIs
Publication statusPublished - 2001 Sep 15

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

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    Keyoung, H. M., Roy, N. S., Benraiss, A., Louissaint, A., Suzuki, A., Hashimoto, M., Rashbaum, W. K., Okano, H., & Goldman, S. A. (2001). High-yield selection and extraction of two promoter-defined phenotypes of neural stem cells from the fetal human brain. Nature Biotechnology, 19(9), 843-850. https://doi.org/10.1038/nbt0901-843