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Engraftment of sorted/expanded human central nervous system stem cells from fetal brain
Author(s) -
Tamaki Stanley,
Eckert Karl,
He Dongping,
Sutton Richard,
Doshe Monika,
Jain Gitanjali,
Tushinski Robert,
Reitsma Michael,
Harris Brent,
Tsukamoto Ann,
Gage Fred,
Weissman Irving,
Uchida Nobuko
Publication year - 2002
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.10412
Subject(s) - neurosphere , biology , stem cell , neural stem cell , microbiology and biotechnology , cd90 , glial fibrillary acidic protein , cd34 , immunology , adult stem cell , cellular differentiation , immunohistochemistry , biochemistry , gene
Direct isolation of human central nervous system stem cells (CNS‐SC) based on cell surface markers yields a highly purified stem cell population that can extensively expand in vitro and exhibit multilineage differentiation potential both in vitro and in vivo. The CNS‐SC were isolated from fetal brain tissue using the cell surface markers CD133 + , CD34 – , CD45 – , and CD24 –/lo (CD133 + cells). Fluorescence‐activated cell sorted (FACS) CD133 + cells continue to expand exponentially as neurospheres while retaining multipotential differentiation capacity for >10 passages. CD133 – , CD34 – , and CD45 – sorted cells (∼95% of total fetal brain tissue) fail to initiate neurospheres. Neurosphere cells transplanted into neonatal immunodeficient NOD‐SCID mice proliferated, migrated, and differentiated in a site‐specific manner. However, it has been difficult to evaluate human cell engraftment, because many of the available monoclonal antibodies against neural cells (β‐tubulin III and glial fibrillary acidic protein) are not species specific. To trace the progeny of human cells after transplantation, CD133 + ‐derived neurosphere cells were transduced with lentiviral vectors containing enhanced green fluorescent protein (eGFP) expressed downstream of the phosphoglycerate kinase promoter. After transduction, GFP + cells were enriched by FACS, expanded, and transplanted into the lateral ventricular space of neonatal immunodeficient NOD‐SCID brain. The progeny of transplanted cells were detected by either GFP fluorescence or antibody against GFP. GFP + cells were present in the subventricular zone‐rostral migrating stream, olfactory bulb, and hippocampus as well as nonneurogenic sites, such as cerebellum, cerebral cortex, and striatum. Antibody against GFP revealed that some of the cells displayed differentiating dendrites and processes with neurons or glia cells. Thus, marking human CNS‐SC with reporter genes introduced by lentiviral vectors is a useful tool with which to characterize migration and differentiation of human cells in this mouse transplantation model. © 2002 Wiley‐Liss, Inc.