[Isolation and culture of neural stem cell from rat olfactory epithelium].
OBJECTIVE
To isolate and culture neural stem cell (NSC) from rat olfactory epithelium and investigate the characteristics of its proliferation and differentiation.


METHODS
NSC from postnatal three-day (P3) and zinc sulfate in situ injured adult rat olfactory epithelium was cultured with DMEM/F12 (1 : 1) containing 10% heat-inactivated fetal bovine serum. NSC and specific neural cells differentiated from NSC were identified with indirect immunofluorescence. Growth curve of NSC from P3 and adult rat and the effect of growth factors were measured using MTT method.


RESULTS
Neurosphere-forming cells were isolated and collected with diluted purification method from P3 and adult rat olfactory epithelium. These neurosphere-forming cells were nestin immuno-positive and cytokeratin immuno-negative, and could differentiated into neuronal specific enolase (NSE) immuno-positive neurons and glial fibrillary acidic protein (GFAP) immuno-positive astrocytes when transferred in MEM. The viability of NSC had no significant difference between P3 and adult rat (P > 0.05). The forming rate of neurosphere of P3 and adult rat olfactory epithelium was 0.05% - 0.10%. Fibroblast growth factor-basic could significantly promote the proliferation of NSC from epithelium, while epidermal growth factor showed no effect on the proliferation of the NSC.


CONCLUSIONS
NSC with self-renewal capacity and multipotential differentiation can be isolated and cultured from P3 and adult rat olfactory epithelium.
Chen
Fuquan
F
Department of Otorhinolaryngology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. chenfq@fmmu.edu.cn
Qiu
Jianhua
J
Wang
Jinling
J
Liu
Shunli
S
Mi
Wenjuan
W
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D005260:Female; D058953:Neural Stem Cells / Q000166:cytology*; D009831:Olfactory Mucosa / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley