【摘要】 目的:观察不同剂量神经节苷脂(GM-1)对神经干细胞增殖和分化的影响。方法:从人胚胎脑组织分离出神经干细胞,培养于含bFGF(阳性对照组)和B27(阴性对照组)的DMEM/F12细胞培养液中,实验组培养液中分别加入0.335μg/L,3.35μg/L及33.5μg/L的GM-1,用MTT比色分析法测定细胞增殖能力。用含体积分数3％血清的DMEM/F12培养液诱导细胞分化,每间隔6 h于倒置显微镜下观察细胞分化情况。结果:5 d、10 d MTT比色显示,GM-1(33.5μg/L)组与阳性对照组比较,P均>0.05,与阴性对照组比较,P均<0.05;GM-1(0.335μg/L)组、GM-1(3.35μg/L)组与阴性对照组比较,P均>0.05。分化实验发现,接种6 h后,体积分数3％血清+DMEM/F12组神经球周边可见明显的细胞突起,呈放射状向周围伸出,而3个实验组和阴性对照组的细胞突起短且细小,随着培养时间的延长,各组的细胞突起均逐渐伸长,3个实验组分化能力低于对照组,与阴性对照组之间无明显差异。结论:GM-1能够维持神经干细胞的原始神经状态,促进神经干细胞的增殖,对神经干细胞无明显的诱导分化作用。更多还原

【Abstract】 Aim: To observe the effects of different dose of GM-1 on the growth and differentiation of neural stem cells. Methods; The neural stem cells were isolated from human embryonic brains and cultured in serum-free medium-DMEM/F12 with bFGP’ and B27 ,Different dosage of GM-1 was added into the different experimental group,and the proliferation ability of the cells were measured by MTT methods. The differentiation of the cells was induced in the medium-DMEM/F12 with 3% serum and observed through inverted phase-contrast microscope every 6 hours. Results: According to t test,the 5 d and 10 d MTT showed as follows: there was no significant difference between bFGF(20 μg/L) + B27 + DMEM/F12 and GM-1(33.5 μg/L) + B27 + DMEM/F12, however,the significant difference lay in GM-1 (33. 5 μg/ml) + B27 + DMEM/F12 and B27 + DMEM/F12, there was no significant differences among GM-1 (0. 335 μg/L) + B27 + DMEM/F12,GM-1(3.35 μg/L) + B27 + DMEM/F12 and B27 + DMEM/F12. The differential experiments showed that: the neurites from the neuralspheres of (3% )serum + DMEM/F12 group stuck in a radial way after the neural stem cells being replanted in the 24-well plate coated with poly-lysine for six hours while the neurites of the three experimental groups and the negative-control group were short and thin, the neurites of each group developed little by little with the time gone by,however, the neurites of the three experimental groups and the negative-control group were much less than those of positive-control group. Conclusion: GM-1 is able to maintain the primitive situation of neural stem cell and has effects on its growth, however,it has no obvious influence on its differentiation.更多还原