Morphological and biomolecular characterization of the neonatal olfactory bulb ensheathing cell line.
Cell transplantation therapy has raised a great interest in the perspective of its employment for nerve tissue repair. Among the various cell populations proposed, olfactory ensheathing glial cells have raised great interest over recent years, especially in the perspective of their employment for neural repair because of their homing capacity in both central and peripheral nervous system. This paper is aimed to provide an in vitro characterization of the NOBEC (neonatal olfactory bulb ensheathing cell) line that was obtained from primary cells dissociated from rat neonatal olfactory bulb (OB) and immortalized by retroviral transduction of SV40 large T antigen. Light and electron microscopy investigation showed that NOBECs are a homogeneous cell population both at structural and ultrastructural level. RT-PCR, Western blotting and immunocytochemistry showed that NOBECs express the glial markers S100, GFAP (Glial Fibrillar Acid Protein) and p75NGFR as well as NRG1 (neuregulin-1) and ErbB1-2-3 receptors; while they are negative for ErbB4. Yet, NOBECs exhibit a high proliferation and migration basal activity and can be transducted with vectors carrying GFP (green fluorescent protein) and NRG1 cDNA. Functional stimulation by means of NRG1-III-beta3 overexpression through viral transduction induced a significant increase in cell proliferation rate while it had no effect on cell migration. Altogether, these results show that NOBEC cell line retain glial features both morphologically and functionally, responding to the NRG1/ErbB-mediated gliotrophic stimulus, and represents thus a good tool for in vitro assays of glial cell manipulation and for in vivo experimental studies of glial cell transplantation in the central and peripheral nervous system.
Audisio
Chiara
C
Department of Animal and Human Biology, University of Torino, Torino, Italy.
Raimondo
Stefania
S
Nicolino
Silvia
S
Gambarotta
Giovanna
G
Di Scipio
Federica
F
Macrì
Loredana
L
Montarolo
Francesca
F
Giacobini-Robecchi
Maria Giuseppina
MG
Porporato
Paolo
P
Filigheddu
Nicoletta
N
Graziani
Andrea
A
Geuna
Stefano
S
Perroteau
Isabelle
I
D000818:Animals; D000831:Animals, Newborn; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D015153:Blotting, Western; D016380:Brain Tissue Transplantation / Q000379:methods*; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism; D020890:Neuregulin-1 / Q000235:genetics; D009457:Neuroglia / Q000378:metabolism* / Q000637:transplantation / Q000648:ultrastructure*; D009830:Olfactory Bulb / Q000378:metabolism* / Q000637:transplantation / Q000648:ultrastructure*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic / Q000379:methods*; D014780:Viruses / Q000235:genetics
D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D020890:Neuregulin-1; C490580:Nrg1 protein, rat; D049452:Green Fluorescent Proteins