Abstract
BackgroundProgenitor cells isolated from adult brain tissue are important tools for experimental studies of remyelination. Cells harvested from neurogenic regions in the adult brain such as the subependymal zone have demonstrated remyelination potential. Multipotent cells from the progenitor fraction have been isolated from the adult olfactory bulb (OB) but their potential to remyelinate has not been studied.Methodology/Principal FindingsWe used the buoyant density gradient centrifugation method to isolate the progenitor fraction and harvest self-renewing multipotent neural cells grown in monolayers from the adult green-fluorescent protein (GFP) transgenic rat OB. OB tissue was mechanically and chemically dissociated and the resultant cell suspension fractionated on a Percoll gradient. The progenitor fraction was isolated and these cells were plated in growth media with serum for 24 hrs. Cells were then propagated in N2 supplemented serum-free media containing b-FGF. Cells at passage 4 (P4) were introduced into a demyelinated spinal cord lesion. The GFP+ cells survived and integrated into the lesion, and extensive remyelination was observed in plastic sections. Immunohistochemistry revealed GFP+ cells in the spinal cord to be glial fibrillary acidic protein (GFAP), neuronal nuclei (NeuN), and neurofilament negative. The GFP+ cells were found among primarily P0+ myelin profiles, although some myelin basic protein (MBP) profiles were present. Immuno-electron microscopy for GFP revealed GFP+ cell bodies adjacent to and surrounding peripheral-type myelin rings.Conclusions/SignificanceWe report that neural cells from the progenitor fraction of the adult rat OB grown in monolayers can be expanded for several passages in culture and that upon transplantation into a demyelinated spinal cord lesion provide extensive remyelination without ectopic neuronal differentiation.
Highlights
We have shown in our previous work [1,2] that neural progenitor cultures from a number of different adult brain regions converge to a similar morphology and potential in vitro
Conclusions/Significance: We report that neural cells from the progenitor fraction of the adult rat olfactory bulb (OB) grown in monolayers can be expanded for several passages in culture and that upon transplantation into a demyelinated spinal cord lesion provide extensive remyelination without ectopic neuronal differentiation
Grafted green-fluorescent protein (GFP)+ cells produced robust, extensive remyelination of the spinal cord with a largely peripheral pattern of myelin. These results indicate that cells from the OB progenitor fraction are capable of remyelinating axons in the central nervous system (CNS) with a predominantly peripheral Schwann-like pattern of myelination, similar to that reported for other central neural progenitors, and indicate that the OB may provide a rich source of cells for remyelination
Summary
We have shown in our previous work [1,2] that neural progenitor cultures from a number of different adult brain regions converge to a similar morphology and potential in vitro. In culture, differentiated progenitors isolated from the adult hippocampus (an alar plate derivative) are able to produce the classic hypothalamic (basal plate derivative) neuroendocrine peptides oxytocin, vasopressin, thyrotropin-releasing hormone, growth hormone-releasing hormone, somatostatin, corticotropin-releasing hormone, and gonadotropin-releasing hormone (GnRH; [2]). This is noteworthy in the case of. The neurogenic hippocampus and subependymal zone of the lateral ventricles [7,8,9: for a review see 10] were the initial targets for progenitor isolation, but subsequent work has uncovered progenitors in other brain regions including the isocortex, optic nerve, hypothalamus; each of these areas has yielded self-renewing cultures capable in the presence of b-FGF of generating cells of all three neural lineages [1,2]. Multipotent cells from the progenitor fraction have been isolated from the adult olfactory bulb (OB) but their potential to remyelinate has not been studied
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