摘要
目的 :探讨成年小鼠大脑室下区神经干细胞(neural stem cells,NSCs)体外分离培养和鉴定的方法 ,测定体外培养的神经干细胞多不饱和脂肪酸n6/n3的比例和二十二碳六烯酸(docosahexaenoic acid,DHA)含量。方法:取8~10周龄的C57BL/6小鼠室下区细胞,进行体外悬浮培养,观察NSCs形成情况,免疫荧光鉴定NSCs干细胞特性及体外分化能力,气相色谱鉴定NSCs的多不饱和脂肪酸含量中n6/n3比例及DHA含量。结果:体外培养的细胞可以增殖,并可以连续传代;免疫荧光显示神经干细胞呈抗巢蛋白阳性,神经干细胞在分化培养基培养时可分化为NeuN、TuJ1、GFAP、O4阳性细胞;神经干细胞的多不饱和脂肪酸含量较鼠尾高。结论:采用无血清培养基成功获得成年小鼠室下区的神经干细胞,具有增殖、自我更新和分化的能力;神经干细胞的n3多不饱和脂肪酸含量明显高于鼠尾中的含量,且DHA多不饱和脂肪酸占较高比例。
Objective：To in vestigate the method of culturing neural stem cell s（NSCs） in vitro and measurement of n6/n3 ratios and DHA contents in these NSCs cultured from subventricular zone in adult mice. Methods：Subventricular zone was isolated from the brains of 8~10 weeks old C57BL/6 mice, and the NSCs were cultured in floating in vitro. Some marker genes of NSCs and differentiated cells were detected by immunofluorescendce; polyunsaturated fatty acids and n6/n3 ratios were measured by gas chromatography. Results：NSCs proliferated continuously in vitro, most cells were Nestin positive. NeuN,TuJ1,GFAP and O4 positive cells were detected by immunofluorescence in differentiated neural cells. Polyunsaturated fatty acids were richer in NSCs. Conclusion：NSCs cultured from subventricular zone in adult mice have proliferation and self-renewal abilities. n3 polyunsaturated fatty acids and DHA in NSCs are significantly higher than those in tails.
出处
《南京医科大学学报（自然科学版）》
CSCD
北大核心
2017年第4期394-398,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81570402)
南京医科大学科技发展基金(2011NJMU256)
