FGF-2,NT-3对海马表皮生长因子反应性神经干细胞分化为神经元的诱导作用

目的:比较碱性成纤维细胞生长因子(FGF-2)和神经营养素3(NT-3)对海马表皮生长因子(EGF)反应性神经干细胞(NSC)分化为神经元的诱导作用。方法:在含EGF的培养体系中分离培养NSC,免疫荧光染色检测NSC特征性标志nestin的表达,Brdu掺入实验检测细胞增殖状态。将NSCs接种在6孔培养板中,分成4组:FGF-2组、NT-3组、FGF-2+NT-3组和对照组。培养10d时,用免疫细胞化学染色和荧光标记技术检测其神经元特异性烯醇化酶(NSE)的表达,并计算阳性染色细胞的百分率。结果:FGF-2和NT-3均可诱导EGF反应性NSC分化为神经元,FGF-2优于NT-3。对照组神经元细胞分化率低,仅为3.70%。联合应用FGF-2和NT-3,可使神经元分化率增加约10倍。结论:FGF-2与NT-3联合作用能更好地促进海马EGF反应性NSC分化为神经元细胞。

Inducing Effect of FGF-2 in Combination with NT-3 on Differentiation of EGF-responsive Neural Stem Cells into Neurons

Objective: To compare the effects of FGF2 and NT-3 on the differentiation of EGF-responsive neural stem cells in cultures.Methods: NSCs were cultured in DMEM/F12 medium containing EGF.Nestin was detected by immunofluorescent labeling,and Brdu was used to show cell stages in division.EGF-responsive NSCs plated into 6 well culture-plates were divided into 4 groups: 20ng/ml FGF-2,40ng/ml NT-3,FGF-2+NT-3 and the control groups.10 days after NSCs were cultured,NSE expression could be detected using immunocytochemical and immunofluorescent staining.The number of NSE positive cells was both counted and the percentage of positive cells was obtained respectively.Results: Most of the cultured cells were nestin-positive and Brdu-positive.Both FGF-2 and NT-3 can induce EGF-responsive NSCs to differentiate into neurons in vitro.The effect of FGF-2 on differentiation is more optimal than that of NT-3.The number of differentiated neurons in NT-3、FGF-2、FGF-2+ NT-3 and the control groups were 10.52%、19.69%、34.10%、3.70% respectively.Combination of NT-3 and FGF-2 could increase differentiation by about 10 times.Conclusion: Combination of FGF-2 and NT-3 in inducing differentiation of NSCs into neurons in vitro is recommended.