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Updated: Jan 26, 2026

Quadruple Immunostaining of the Olfactory Bulb for Visualization of Olfactory Sensory Axon Molecular Identity Codes
Published on: June 5, 2017
[Culture of neural stem cells from rat olfactory bulb]
Fu-quan Chen1, Jian-hua Qiu, Jin-ling Wang
1Department of Otorhinolaryngology, Xijing Hospital, Fourth Military Medical University, Xi'an, China. chenfq@fmmu.edu.cn
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.
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