神经生长因子对神经干细胞分化的影响

Effects of Neural Growth Factors on the Differentiation of Neural Stem Cells.

【目的】探讨不同神经生长因子对神经干细胞(neural stemcells,NSCs)分化特性的影响。【方法】自10～14周人胚胎脑分离培养海马神经干细胞,分别用表皮生长因子(human epidermal growth factor,EGF)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF),或单独用bFGF扩增神经干细胞,取第3代神经干细胞球,用含小剂量bFGF(0.5 ng)的无血清基础培养基诱导分化,体外诱导分化7 d后应用免疫荧光方法检测β_Ⅲ微管蛋白(β_ⅢTubulin,Tuj1)和神经胶质纤维酸性蛋白(glial fibriliary acidic protein,GFAP)表达情况。【结果】不同的生长因子培养的神经干细胞分化特性很不相同,EGF+bFGF培养的NSCs既可分化为神经元,又可分化为星形胶质细胞,其分化比率分别为38.8%和57.3%;bFGF培养的NSCs主要分化为神经元,比率达81.2%,未检测星形胶质细胞。【结论】不同神经生长因子扩增的神经干细胞分化特性很不相同,本实验为神经干细胞分化的研究提供了良好的实验平台。

【Objective】 To explore the effects of different neural growth factors on the differentiation of neural stem cells（NSCs）. 【Methods】 NSCs were separated from hippocampus of 10w to 14w human fetus brains and incubated in serum-free DMEM/F12 medium containing human epidermal growth factor（EGF） and basic fibroblastgrowth factor（bFGF） or bFGF alone respectively.Neurosphere cultures from passage three were induced to differentiate with serum-free media containing 0.5ng bFGF.After 7-day differentiation,expression of β_ⅢTubulin（Tuj1） and glial fibriliary acidic protein（GFAP） was detected by immunofluorescence. 【Results】 The differentiation characteristics of NSCs generated with different growth factors were different： EGF＋bFGF generated NSCs differentiated into both neurons and astrocytes,and the ratio of them were 38.8% and 57.3% respectively.bFGF generated NSCs mostly differentiated into neurons,the ratio of neurons was 81.2%,and few astrocytes were detected. 【Conclusions】 NSCs cultured with different growth factors have completely different differentiation characteristics,this research may provide experimental platform for the study of NSCs differentiation.