title: Isolation and characterization of neural progenitor cells from post-mortem human cortex.
abstract: Post-mortem human brain tissue represents a vast potential source of neural progenitor cells for use in basic research as well as therapeutic applications. Here we describe five human neural progenitor cell cultures derived from cortical tissue harvested from premature infants. Time-lapse videomicrography of the passaged cultures revealed them to be highly dynamic, with high motility and extensive, evanescent intercellular contacts. Karyotyping revealed normal chromosomal complements. Prior to differentiation, most of the cells were nestin, Sox2, vimentin, and/or GFAP positive, and a subpopulation was doublecortin positive. Multilineage potential of these cells was demonstrated after differentiation, with some subpopulations of cells expressing the neuronal markers beta-tubulin, MAP2ab, NeuN, FMRP, and Tau and others expressing the oligodendroglial marker O1. Still other cells expressed the classic glial marker glial fibrillary acidic protein (GFAP). RT-PCR confirmed nestin, SOX2, GFAP, and doublecortin expression and also showed epidermal growth factor receptor and nucleostemin expression during the expansion phase. Flow cytometry showed high levels of the neural stem cell markers CD133, CD44, CD81, CD184, CD90, and CD29. CD133 markedly decreased in high-passage, lineage-restricted cultures. Electrophysiological analysis after differentiation demonstrated that the majority of cells with neuronal morphology expressed voltage-gated sodium and potassium currents. These data suggest that post-mortem human brain tissue is an important source of neural progenitor cells that will be useful for analysis of neural differentiation and for transplantation studies.
authors.affiliation: National Human Neural Stem Cell Resource, Children's Hospital of Orange County Research Institute, Orange, California 92868-3874, USA. pschwartz@choc.org
authors.forename: Philip H
authors.initials: PH
authors.lastname: Schwartz
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Bryant
authors.forename: Tannin J
authors.initials: TJ
authors.lastname: Fuja
authors.forename: Hailing
authors.initials: H
authors.lastname: Su
authors.forename: Diane K
authors.initials: DK
authors.lastname: O'Dowd
authors.forename: Henry
authors.initials: H
authors.lastname: Klassen
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007231:Infant, Newborn; D007621:Karyotyping / Q000379:methods; D008297:Male; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D009474:Neurons / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000473:pathology
chemical_list: D009419:Nerve Tissue Proteins