Primary Culture and Identification of Olfactory Ensheathing Cells from Adult Rat

Objective To explore a simple and inexpensive method or procedure for establishing primary culture and purification of olfactory ensheathing cells(OECs)from the adult rat olfactory bulbs.Methods The OECs were dissociated from the first two outer layers of the olfactory bulbs of the adult Sprague-Dawley rat(2.5 months old),cultured in DMEM/F12 nutrient with 20% foetal calf serum,and purified by the method of combining the different rates of cell attachment with the Arabinosylcytosine(AraC)inhibition of cell.The morphological changes of cultured OECs were observed.On 14th culture day,the OECs in culture were identified by the immunocytochemistry technique to glial fibrillary acidic protein(GFAP)and nerve growth factor receptor p75(NGFRp75),and the purity of the positive cells was calculated.Results The cultured OECs presented three main morphological types:multipolar,bipolar and flat cells.For GFAP positive cells,the plasma and processes were stained while the nuclei were not,but for NGFRp75 cells the nuclei were stained more deeply than the plasma and processes.More than 90% cultured cells were identified to be OECs.Conclusion The high purity OECs can be cultured successfully in vitro by the combined purification method.This method is simple and inexpensive.