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TISSUE-SPECIFIC STEM CELLS

In Vitro–Derived "Neural Stem Cells" Function as Neural Progenitors Without the Capacity for Self-Renewal

Program in Stem Cell Biology and Regenerative Medicine, University of Florida, Gainesville, Florida, USA

Key Words. Green fluorescent protein • Multipotent astrocytic stem cell • Neurosphere • Neural stem cell • Subependymal zone • Transplantation

Correspondence: Gregory P. Marshall II, Ph.D., Program in Stem Cell Biology and Regenerative Medicine, University of Florida, 1600 SW Archer Road, Gainesville, Florida 32611, USA. Telephone: 352-392-0231; Fax: 352-392-0025; e-mail: gpm2{at}ufl.edu

Received May 1, 2005; accepted for publication August 22, 2005.
Hematopoietic stem cells have been defined by their ability to self-renew and successfully reconstitute hematopoiesis throughout the life of a transplant recipient. Neural stem cells (NSCs) are believed to exist in the regenerating regions of the brain in adult mice: the subependymal zone (SEZ) of the lateral ventricles (LVs) and the hippocampal dentate gyrus. Cells from the SEZ can be cultured to generate neurospheres or multipotent astrocytic stem cells (MASCs), both of which demonstrate the stem cell qualities of multipotency and self-renewal in vitro. Whether neurospheres and MASCs possess the true stem cell quality of functional self-renewal in vivo is unknown. The definitive tests for this unique capability are long-term engraftment and serial transplantation. Both neurospheres and MASCs transplanted into the LVs of C57BL/6 mice resulted in short-term engraftment into the recipient brain, with donor-derived migratory neuroblasts visible in the rostral migratory stream and olfactory bulb after transplantation. To test in vivo expansion/self-renewal of the transplanted cells, we attempted to reisolate donor-derived neurospheres and MASCs. Even when rigorous drug selection was used to select for rare events, no donor-derived neurospheres or MASCs could be reisolated. Furthermore, donor-derived migratory neuroblasts were not observed in the rostral migratory stream (RMS) for more than 1 month after transplantation, indicating a transient rather than long-term engraftment. Therefore, in vitro-derived neurospheres and MASCs do not function as NSCs with long-term, self-renewal capabilities in vivo but instead represent short-term neural progenitor cells as defined by an in vivo functional assay.

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