Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone.
Adult humans, like their nonhuman mammalian counterparts, harbor persistent neural progenitor cells in the forebrain ventricular lining. In the absence of adequate surface markers, however, these cells have proven difficult to isolate for study. We have previously identified and selected neural progenitor cells from both the fetal and adult rodent ventricular zone (VZ), by sorting forebrain cells transfected with plasmid DNA encoding the gene for green fluorescent protein driven by the early neuronal promoter for Talpha1 tubulin (P/Talpha1:hGFP). We have now extended this approach by purifying both P/Talpha1:hGFP tubulin-defined neuronal progenitors, as well as potentially less committed E/nestin:hGFP-defined neural progenitor cells, from the adult human VZ. The ventricular wall of the temporal horn of the lateral ventricle was dissected from temporal lobes obtained from four adult patients undergoing therapeutic lobectomy. These samples were dissociated, and the cultured cells transduced with either P/Talpha1:hGFP or E/nestin:EGFP plasmid DNA. A week later, the cells were redissociated, selected via fluorescence-activated cell sorting (FACS) on the basis of neural promoter-driven GFP expression, and replated. The majority of these cells expressed the early neuronal protein betaIII-tubulin upon FACS; within the week thereafter, most matured as morphologically evident neurons that coexpressed betaIII-tubulin and microtubule-associated protein (MAP)-2. Many of these neurons had incorporated bromodeoxyuridine in vitro in the days before FACS, indicating their mitogenesis in vitro. Thus, the use of fluorescent transgenes under the control of early neural promoters permits the enrichment of neuronal progenitor cells from the adult human ventricular zone. The specific acquisition, in both purity and number, of residual neural progenitor cells from the adult human brain may now permit hitherto unfeasible studies of both their biology and practical application.
Roy
N S
NS
Department of Neurology and Neuroscience, Cornell University Medical College, New York, New York 10021, USA.
Benraiss
A
A
Wang
S
S
Fraser
R A
RA
Goodman
R
R
Couldwell
W T
WT
Nedergaard
M
M
Kawaguchi
A
A
Okano
H
H
Goldman
S A
SA
D000328:Adult; D002455:Cell Division; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002552:Cerebral Ventricles / Q000166:cytology*; D002675:Child, Preschool; D004742:Enhancer Elements, Genetic; D005434:Flow Cytometry; D018390:Gene Targeting; D049452:Green Fluorescent Proteins; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D009419:Nerve Tissue Proteins; D064231:Nestin; D009474:Neurons / Q000166:cytology*; D011401:Promoter Regions, Genetic; D013234:Stem Cells / Q000166:cytology*; D014404:Tubulin / Q000235:genetics
D007381:Intermediate Filament Proteins; D008164:Luminescent Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D014404:Tubulin; D049452:Green Fluorescent Proteins