Chen Fu-quan, Qiu Jian-hua, Wang Jin-ling, Liu Shun-li, Mi Wen-juan
Department of Otorhinolaryngology, Xijing Hospital, Fourth Military Medical University, Xi'an, China.
Zhonghua Er Bi Yan Hou Ke Za Zhi. 2004 Nov;39(11):687-90.
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.
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.
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.
NSC with self-renewal capacity and potential multi-differentiation were cultured from P1 and adult rat olfactory bulb.
建立大鼠嗅球神经干细胞（NSC）的体外培养方法，并研究其增殖和分化特性。
分离出生1天（P1）和成年大鼠嗅球的神经干细胞，在含有表皮生长因子（EGF）和成纤维细胞生长因子-碱性（bFGF）的无血清培养基中培养。使用针对神经干细胞（巢蛋白）、神经元（神经元特异性烯醇化酶，NSE）、星形胶质细胞（胶质纤维酸性蛋白，GFAP）和少突胶质细胞（半乳糖脑苷脂，GalC）标志物的抗体，通过免疫细胞化学染色鉴定神经干细胞和从神经干细胞分化而来的特定神经细胞。采用MTT法测定嗅球神经干细胞的生长曲线。
从P1和成年大鼠嗅球中分离培养出巢蛋白免疫阳性的神经干细胞，其可分化为神经元、星形胶质细胞和少突胶质细胞。P1和成年大鼠嗅球神经球的形成率分别约为20%至30%和0.1%。嗅球神经干细胞的增殖依赖于EGF和bFGF，其中EGF对细胞增殖的促进作用强于bFGF。
从P1和成年大鼠嗅球中培养出了具有自我更新能力和多向分化潜能的神经干细胞。
