[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 9000 cultured cells were identified to be OECs.


CONCLUSIONS
The high purity OECs can be cultured successfully in vitro by the combined purification method. This method is simple and inexpensive.
Li
Shao-bo
SB
Department of Histology, Embryology and Neurobiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Zhang
Wei
W
Ding
Yan
Y
Peng
Jin
J
Zhu
Zhen-dong
ZD
Zhou
Xue
X
D000818:Animals; D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods*; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation / Q000191:economics / Q000379:methods*; D048430:Cell Shape; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D009830:Olfactory Bulb / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling
D005904:Glial Fibrillary Acidic Protein