[Culture of neural stem cells from rat olfactory bulb].
OBJECTIVE
To establish the method of in vitro culture of neural stem cells (NSC) from rat olfactory bulb and investigate the characteristics of its proliferation and differentiation.


METHODS
NSC from postnatal one-day (P1) and adult rat olfactory bulb were isolated and cultured in serum free media with epidermal growth factor (EGF) and fibroblast growth factor-basic (bFGF). Antibodies against NSC (nestin), neuronal (neuronal specific enolase, NSE), astrocytic (glial fibrillary acidic protein, GFAP) and oligodendrocytic (galactocerebroside, GalC) markers were used to identify NSC and specific neural cells differentiated from NSC with immunocytochemical staining. Growth curve of olfactory bulb NSC was measured using MTT method.


RESULTS
Nestin immuno-positive NSC were isolated and cultured from P1 and adult rat olfactory bulb which could differentiate into neurons, astrocytes and oligodendrocytes. The forming rates of neurosphere from P1 and adult rat olfactory bulb were 20% approximately 30% and 0.1% respectively. The proliferation of olfactory bulb NSC depended on EGF and bFGF, in which EGF increased proliferation of cells stronger than bFGF.


CONCLUSIONS
NSC with self-renewal capacity and potential multi-differentiation were cultured from P1 and adult rat olfactory bulb.
Chen
Fu-quan
FQ
Department of Otorhinolaryngology, Xijing Hospital, Fourth Military Medical University, Xi'an, China. chenfq@fmmu.edu.cn
Qiu
Jian-hua
JH
Wang
Jin-ling
JL
Liu
Shun-li
SL
Mi
Wen-juan
WJ
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D008297:Male; D058953:Neural Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology; D009830:Olfactory Bulb / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley