Culture, Purification And Biological Characteristics Of Olfactory Ensheathing Cells From The Adult Rat Olfactory Bulbs And Olfactory Mucosa

Part 1: Isolation,Culture, Purification and Biological Characteristics of Olfactory Ensheathing Cells from the adult rat Olfactory bulbsObjcctive: To culture and purify the olfactory ensheathing cells from the adult rat olfactory bulb(sOB-OECs) in vitro,and to investigate the morphological changes and the immunocytochemical properties of the cultured OB-OECs.Methods: The OB-OECs were dissociated from the adult rat olfactory bulbs. The primarily cultured OB-OECs were purified based on the differing rates of attachment of the various harvested cell types and/or anti-Thy 1.1-mediated complement lysis. The morphological changes of the cultured OB-OECs were observed under a phase contrast microscope at different culture time. The OB-OECs were identified by GFAP,NGFRp75 and S100 immunocytochemistry. The purity of the OB-OECs was evaluated according to the percentage of NGFRp75 immunostaining cells.Results: The OB-OECs displayed a very characteristic morphological appearance. Most of OB-OECs were bipolar or tripolar with long and thin processes. In the unpurified group the rapidly proliferating fibroblasts were in the majority after 10 days in culture. The OB-OECs puritification by the method based on the differing rates of attachment of the various harvested cell types with anti-Thy 1.1-mediated complement lysis was more efficient than the others. bFGF could facilitate OB-OECs growth,and could not affect the proportion of positive of OB-OECs.