TY - JOUR
T1 - Morphological and biomolecular characterization of the neonatal olfactory bulb ensheathing cell line
AU - Audisio, Chiara
AU - Raimondo, Stefania
AU - Nicolino, Silvia
AU - Gambarotta, Giovanna
AU - Di Scipio, Federica
AU - Macrì, Loredana
AU - Montarolo, Francesca
AU - Giacobini-Robecchi, Maria Giuseppina
AU - Porporato, Paolo
AU - Filigheddu, Nicoletta
AU - Graziani, Andrea
AU - Geuna, Stefano
AU - Perroteau, Isabelle
N1 - Funding Information:
The authors wish to thank Prof. Luigi Naldini for providing the MA1 lentiviral vector, Prof. Mauro Giacca for providing the pAAV-MCS vector. The authors also wish to thank Prof. Roberto Piva and Dr. Chiara Ambrogio for providing facilities and expert assistance for lentiviral vector manipulation. This work was supported by grants from the Regione Piemonte (“Bando Ricerca Sanitaria Finalizzata”) , the Ministero Italiano dell’Università e della Ricerca (MIUR) , FIRB fund (code: RBAU01BJ95), PRIN2005 fund (code: 2005057088), the University of Turin (ex-60% fund), and the Compagnia di San Paolo (“Bando Programma Neuroscienze”). Dr. Stefania Raimondo is recipient of a PostDoc grant partially funded by the Regione Piemonte (“Azione Contenimento del Brain Drain”).
PY - 2009/12/15
Y1 - 2009/12/15
N2 - 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-β3 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.
AB - 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-β3 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.
KW - ErbB receptors
KW - Gene transfer
KW - Neuregulin-1
KW - Olfactory glia
UR - https://www.scopus.com/pages/publications/70350732786
U2 - 10.1016/j.jneumeth.2009.09.021
DO - 10.1016/j.jneumeth.2009.09.021
M3 - Article
SN - 0165-0270
VL - 185
SP - 89
EP - 98
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
IS - 1
ER -