Culture and morphologic characteristics of olfactory ensheathing cells isolated from young adult GFP rat

Abstract

Objective To establish methods for isolating and culturing the olfactory ensheathing cells (OECs) from young adult GIT rat to lay the foundation for repair of the spinal cord injury.Methods The OECs were dissociated from the first outer layer of the olfactory bulb of the young adult GFP rat (2.5 months old) under anatomical microscope; enzymatic digestion was performed on these cells and then they were inoculated into DMEM/F12 with 20% fetal calf serum. The morphology of OECs was regularly observed and photographied under light and electronic microscope. On the 10th culture day, the OECs were identified by the immunocytochemistry staining of S-100 and NGFRp75 and the purity (the positive rate) was calculated. Results The OECs showed strong green fluorescence under fluorescent microscope and presented morphological types of bipolar and bearing multipolar. More than 95% of the cultured cells were identified to be OECs, which expressed S-100 and NGFR p75. The cell structure revealed by electron microscope was much accordance with that by light microscope. Conclusion This method is easy to perform and high purity of GFP-OECs can be harvested with the same morphological characteristics and biological activity as general OECs. Therefore, OECs derived from the GFP transgenic rat can be effective tool cells, and widely used in studying the role of OECs in the repair of spinal cord injury Key words: Rat; Olfectory ensheathing cell; Cell culture