Schwann cells (SC) play essential roles in nerve regeneration by the secretion of trophic support molecules and the establishment of a supportive growth guide regrowing axons toward their origin. In the present study, we compared primary peripheral Schwann cells and glial cells from olfactory system (Olfactory Ensheathing Cells-OECs) with an immortalized cell line - the Neonatal Olfactory Bulb Ensheathing Cells (NOBEC) - and with a Schwannoma cell line (RT4-D6P2T) to identify a good cell model for in vitro assays of glial cell manipulation and for in vivo experimental studies. This aspect is particularly important because preparation of primary cultures has several disadvantages such as a limited cell number, a possible contamination with fibroblasts, astrocytes or other cells, and short time survival in culture. To this purpose, we evaluated - by quantitative real time-PCR - the mRNA expression of different neuregulin-1 isoforms, of ErbB receptors and of the principal glial markers, GFAP, P75 and S100. Protein expression was also confirmed by Western blotting. Our results show interesting differences in gene expression between the cell lines (NOBEC and RT4-D6P2T) and primary cell cultures (OECs and SC).
Differential expression of NRG 1 isoforms, ErbBs and glial genes in Schwann cells, Olfactory Ensheathing Cells, Neonatal Olfactory Bulb Ensheathing Cells and RT4-D6P2T
GIOVANNELLI, ALESSIA;GAMBAROTTA, Giovanna;GEUNA, Stefano;PERROTEAU, Isabelle
2012-01-01
Abstract
Schwann cells (SC) play essential roles in nerve regeneration by the secretion of trophic support molecules and the establishment of a supportive growth guide regrowing axons toward their origin. In the present study, we compared primary peripheral Schwann cells and glial cells from olfactory system (Olfactory Ensheathing Cells-OECs) with an immortalized cell line - the Neonatal Olfactory Bulb Ensheathing Cells (NOBEC) - and with a Schwannoma cell line (RT4-D6P2T) to identify a good cell model for in vitro assays of glial cell manipulation and for in vivo experimental studies. This aspect is particularly important because preparation of primary cultures has several disadvantages such as a limited cell number, a possible contamination with fibroblasts, astrocytes or other cells, and short time survival in culture. To this purpose, we evaluated - by quantitative real time-PCR - the mRNA expression of different neuregulin-1 isoforms, of ErbB receptors and of the principal glial markers, GFAP, P75 and S100. Protein expression was also confirmed by Western blotting. Our results show interesting differences in gene expression between the cell lines (NOBEC and RT4-D6P2T) and primary cell cultures (OECs and SC).I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.