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Article
Nature Biotechnology  19, 843 - 850 (2001)
doi:10.1038/nbt0901-843

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

H. Michael Keyoung1, 7, Neeta S. Roy1, 7, Abdellatif Benraiss1, Abner Louissaint Jr.1, Akira Suzuki3, 4, Mitsuhiro Hashimoto5, William K. Rashbaum2, Hideyuki Okano3, 4, 6 & Steven A. Goldman1

1  Department of Neurology and Neuroscience, Cornell University Medical College and New York Presbyterian Hospital, New York, NY 10021.

2  Department of Obstetrics and Gynecology, Cornell University Medical College and New York Presbyterian Hospital, New York, NY 10021.

3  Department of Neuroanatomy, Osaka University School of Medicine, Suita, Osaka 566-0871, Japan.

4  Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation (JST), Suita, Osaka 565-0871, Japan.

5  Laboratory for Developmental Neurobiology, RIKEN BSI, Wako, Saitama 351-0198, Japan.

6  Department of Physiology, Keio University School of Medicine, Shinjuku-ku 160-8582, Tokyo, Japan.

7  These two authors contributed equally to this work.

Correspondence should be addressed to Steven A. Goldman sgoldm@mail.med.cornell.edu
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 musashi1) 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.

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Nature Biotechnology
ISSN: 1087-0156
EISSN: 1546-1696
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