胚胎大鼠嗅神经干细胞的培养及分化特性

目的建立胚胎大鼠嗅神经干细胞(NSCs)体外培养方法,研究其增殖和分化特性.方法采用添加丝裂原的无血清培养基分离、培养胚胎14 d(E14)大鼠嗅球NSCs,应用免疫细胞化学方法鉴定培养的NSCs及自然分化为特异性神经细胞的类型,测定NSCs的生长曲线.结果从E14大鼠嗅球分离、培养出表达nestin,并能分化为神经元、星形胶质细胞和少突胶质细胞的NSCs.嗅NSCs的增殖依赖表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF),其中EGF的促分裂增殖作用明显优于bFGF.结论从E14大鼠嗅球培养出具有自我增殖和多向分化潜能的NSCs.

Culture and differentiation of olfactory neural stem cells from embryonic rat

Objective To establish the method of culturing neural stem cells (NSCs) in vitro from embryonic rat olfactory bulb and investigate their proliferation and differentiation.Methods NSCs from embryonic 14 days' (E14) rat olfactory bulb were isolated and cultured in serum free media with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). Antibodies against nestin, neuronal(neuronal specific enolase, NSE), astrocytic(glial fibrillary acidic protein, GFAP) and oligodendrocytic( galactose-cerebroside, GalC) markers were used to identify NSCs and specific differentiated cells from NSCs with immunocytochemical staining. Growth curve of NSCs was measured.Results Nestin immuno-positive NSCs were isolated and cultured from E14 rat olfactory bulb which could differentiate into neurons, astrocytes and oligodendrocytes. The proliferation of olfactory NSCs depended on EGF and bFGF, and EGF had a stronger effect on the proliferation of cells.Conclusion NSCs with self-renewal capacity and multi-differentiation potential can be cultured from E14 rat olfactory bulb.