In the peripheral nervous system (PNS) Schwann cells (SCs) envelop axons forming myelin, enabling efficient electrical conduction and protection from the extracellular environment. They play essential roles after peripheral nerve injuries by the secretion of trophic support molecules and establish a supportive growth guide for regrowing axons toward their origin. In vivo and in vitro studies are useful to understand the mechanisms involved in myelination and nerve regeneration. However, preparation of in vitro 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 cope with all these problems, immortalized cell lines can represent a good alternative, at least for experimental animal studies. However, immortalization can lead to subtle alteration of cell properties and functional activity and thus in vitro morphological and functional characterization of a cell line is an important step. Therefore, 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. 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 differences in gene expression between the cell lines (NOBEC and RT4-D6P2T) and primary cell cultures (OECs and SC).
Compared expression of Neuregulin-1/ErbBs and glial genes among primary and immortalized cell lines of Schwann cells and Olfactory Ensheathing Cells
GIOVANNELLI, ALESSIA;GAMBAROTTA, Giovanna;GEUNA, Stefano;PERROTEAU, Isabelle
2012-01-01
Abstract
In the peripheral nervous system (PNS) Schwann cells (SCs) envelop axons forming myelin, enabling efficient electrical conduction and protection from the extracellular environment. They play essential roles after peripheral nerve injuries by the secretion of trophic support molecules and establish a supportive growth guide for regrowing axons toward their origin. In vivo and in vitro studies are useful to understand the mechanisms involved in myelination and nerve regeneration. However, preparation of in vitro 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 cope with all these problems, immortalized cell lines can represent a good alternative, at least for experimental animal studies. However, immortalization can lead to subtle alteration of cell properties and functional activity and thus in vitro morphological and functional characterization of a cell line is an important step. Therefore, 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. 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 differences in gene expression between the cell lines (NOBEC and RT4-D6P2T) and primary cell cultures (OECs and SC).
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