中文摘要：

目的观察血管内皮细胞生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）联合应用对胎鼠大脑皮质神经干细胞（NSCs）分化的影响。方法取孕14dSD大鼠,收集胎鼠大脑皮质NSCs,采用无血清培养基传代培养,取P3代细胞,分别加入200μg/LVEGF和（或）20μg/LbFGF进行诱导分化,并设对照组（不加VEGF和bFGF）,7d后采用免疫细胞化学方法检测巢蛋白（Nestin）、β微管蛋白-Ⅲ（β-tubulin-Ⅲ）和胶质纤维酸性蛋白（GFAP）的表达情况。结果从胎鼠大脑皮质中分离的细胞在体外悬浮生长,形成致密的球形细胞团,传代后能形成新的细胞球,免疫细胞化学染色显示细胞球nestin表达阳性。诱导分化后的细胞部分表达β-tubulin-Ⅲ,部分表达GFAP。与对照组比较,VEGF和（或）bFGF均可使NSCs向神经元方向分化的比例增加,其中VEGF＋bFGF联合作用后NSCs的β-tubulin-Ⅲ阳性率（80.3%）最高,GFAP阳性率（18.6%）最低。bFGF作用后NSCs的β-tubulin-Ⅲ阳性率（60.4%）和GFAP阳性率（38.5%）与VEGF作用后（分别为60.3%、38.7%）比较无统计学差异（P〉0.05）。结论VEGF和bFGF均可促进NSCs向神经元分化,且二者具有协同效应。

英文摘要：

Objective To investigate the joint action of vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF） on differentiation of neural stem cells （NSCs） isolated from cortex of rats embryos.Methods The SD rats pregnant for 14 days were selected,and the NSCs of cortex of rat embryos were isolated and cultured in serum-free medium.The third generation cells were treated with VEGF （200μg/L） and/or bFGF （20μg/L）,and the cells of control group were cultured with neither VEGF nor bFGF.The expressions of nestin （neuroectodermal marker）,β-tubulin-Ⅲ （neuron marker） and glial fibrillary acidic protein （GFAP,astrocyte marker） in NSCs were determined by immunocytochemistry 7 days after cultivation.Results The primarily cultured NSCs isolated from embryonic rat cortex grew in a floating state in vitro and formed compact cell masses,and the passage cells could form new cell masses.Most of the primary cells and passage cells expressed nestin.Some differentiated cells expressed β-tubulin-Ⅲ while some others expressed GFAP.Compared with the control group,VEGF and/or bFGF could elevate the proportion of differentiation of NSCs to neurons.The proportion of β-tubulin-Ⅲ positive cells （80.3%） was the highest,while the proportion of GFAP positive cells （18.6%） was the lowest in VEGF＋bFGF group compared with those in other groups.The proportion of β-tubulin-Ⅲ positive cells （60.4%） and GFAP positive cells （38.5%） in bFGF group showed no significant difference compared with those in VEGF group （60.3% and 38.7%,respectively,P0.05）.Conclusion Both VEGF and bFGF can promote the differentiation of NSCs to neurons,and they present a synergistic action.