钙及钙调蛋白依赖性激酶在神经元缺氧性损伤中的作用及其细胞内机理研究

发布时间：2018-06-10 09:10

  本文选题：钙调 + 蛋白　； 参考：《四川大学》2006年博士论文

【摘要】：第一部分 胚鼠大脑皮质神经元的体外培养、鉴定及神经元缺氧模型的建立与评价 目的：建立胚鼠脑皮质神经元培养体系和建立神经元缺氧培养模型，并对神经元缺氧模型进行形态学评价。 方法： 1．改良目前被广泛使用的方法，采取联合使用0.05％胰酶及10IU／mlDNase Ⅰ获取神经元单细胞悬液，以加入2％B27，5％胎牛血清，1％Glutamax的Neurobasal Medium作为接种及维持培养基，接种48小时加入阿糖胞苷抑制胶质细胞生长，综合使用上述方法建立体外神经元培养体系； 2．使用Tubulin β Ⅲ单克隆抗体标记神经元，GFAP多克隆抗体标记星型胶质细胞，运用免疫荧光技术检测胚鼠脑皮质神经元纯度；3．使用国产厌氧培养箱充入85％N2、5％CO2、10％H2混合气建立胚鼠脑皮质神经元的缺氧模型；4．倒置相差显微镜下观察缺氧不同时点胚鼠脑皮质神经元，，对神经元缺氧模型进行形态学评价。
[Abstract]:The first part: in vitro culture and identification of neurons in the cerebral cortex of embryonic mice and the establishment and evaluation of anoxic neuron model objective: to establish the culture system of neurons in the cortex of embryonic rat brain and to establish the model of anoxic culture of neurons. Methods: 1. To improve the current widely used method, we used a combination of 0.05% trypsin and 10 IUU / ml DNase I to obtain single cell suspension of neurons, so as to add 2B275% fetal bovine serum to Neurobasal medium of Glutamax as the inoculation and maintenance medium. 48 hours after inoculation, cytarabine was added to inhibit the growth of glial cells, and the in vitro neuronal culture system was established by using the above methods. 2. The astrocytes were labeled with tubulin 尾 鈪

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