成年小鼠神经干细胞培养、鉴定以及多不饱和脂肪酸含量的测定

发布时间：2018-05-26 08:26

本文选题：神经干细胞 + 神经球　；参考：《南京医科大学学报(自然科学版)》2017年04期

【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。
[Abstract]:Objective: to investigate the method of isolation and identification of neural stem cells (NSCs) from subventricular area of adult mice in vitro, and to determine the proportion of polyunsaturated fatty acids (n6/n3) and the content of 22 carbohexaenoic acid (DHA) in cultured neural stem cells (NSCs). Methods: the cells in the subventricular region of C57BL/6 mice aged 8 ~ 10 weeks were cultured in vitro, the formation of NSCs was observed, and the characteristics and differentiation ability of NSCs stem cells were identified by immunofluorescence. The proportion of n6/n3 and the content of DHA in polyunsaturated fatty acids of NSCs were identified by gas chromatography. Results: the cells cultured in vitro could proliferate and could be subcultured continuously. Immunofluorescence showed that the neural stem cells were positive for anti-nestin, and the neural stem cells could be differentiated into Neun TuJ1 GFAPO4 positive cells when cultured in the differentiation medium. The content of polyunsaturated fatty acids in neural stem cells was higher than that in rat tail. Conclusion: the neural stem cells in the subventricular region of adult mice were successfully obtained by serum-free medium, which had the ability of proliferation, self-renewal and differentiation, and the content of n3 polyunsaturated fatty acids in neural stem cells was significantly higher than that in the tail of mice. And DHA polyunsaturated fatty acids accounted for a high proportion.
【作者单位】：南京医科大学江苏省异种移植重点实验室;南京医科大学附属淮安市第一人民医院病理科;
【基金】：国家自然科学基金(81570402) 南京医科大学科技发展基金(2011NJMU256)
【分类号】：R338

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