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Isolation and characterization of neural progenitor cells from post‐mortem human cortex
Author(s) -
Schwartz Philip H.,
Bryant Peter J.,
Fuja Tannin J.,
Su Hailing,
O'Dowd Diane K.,
Klassen Henry
Publication year - 2003
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.10854
Subject(s) - biology , sox2 , nestin , neural stem cell , glial fibrillary acidic protein , neun , doublecortin , neurosphere , microbiology and biotechnology , progenitor cell , stem cell marker , cd90 , stem cell , cellular differentiation , adult stem cell , immunology , neuroscience , cd34 , embryonic stem cell , immunohistochemistry , genetics , central nervous system , dentate gyrus , gene
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 β‐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. © 2003 Wiley‐Liss, Inc.