TY - JOUR
T1 - High-yield selection and extraction of two promoter-defined phenotypes of neural stem cells from the fetal human brain
AU - Keyoung, H. Michael
AU - Roy, Neeta S.
AU - Benraiss, Abdellatif
AU - Louissaint, Abner
AU - Suzuki, Akira
AU - Hashimoto, Mitsuhiro
AU - Rashbaum, William K.
AU - Okano, Hideyuki
AU - Goldman, Steven A.
N1 - Funding Information:
Acknowledgments Supported by Project ALS, the Human Frontiers Scientific Program, the National Multiple Sclerosis Society, and the Mathers Charitable Foundation. We thank Drs. Theo Palmer and Fred Gage for pNIT-EGFP plasmid and retrovirus, Dr. James Goldman for advice on its use, Drs. Rebecca Baergen and Brad Poulis for assistance in identifying appropriate samples, and Drs. Melissa Carpenter, Kazunobu Sawamoto and Katsuhiko Mikoshiba for valuable discussions, Testu Yoshida for assistance in the preparation of E/nestin:EGFP adenovirus, and Drs. Neil Hackett and Erik Falck-Pedersen for pJM17 and pAdCMV-HSgD, respectively.
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0034869475&partnerID=8YFLogxK
U2 - 10.1038/nbt0901-843
DO - 10.1038/nbt0901-843
M3 - Article
C2 - 11533643
AN - SCOPUS:0034869475
SN - 1087-0156
VL - 19
SP - 843
EP - 850
JO - Nature Biotechnology
JF - Nature Biotechnology
IS - 9
ER -