本文選題：鈣調(diào) + 蛋白　； 參考：《四川大學》2006年博士論文

【摘要】：第一部分 胚鼠大腦皮質(zhì)神經(jīng)元的體外培養(yǎng)、鑒定及神經(jīng)元缺氧模型的建立與評價 目的：建立胚鼠腦皮質(zhì)神經(jīng)元培養(yǎng)體系和建立神經(jīng)元缺氧培養(yǎng)模型，并對神經(jīng)元缺氧模型進行形態(tài)學評價。 方法： 1．改良目前被廣泛使用的方法，采取聯(lián)合使用0.05％胰酶及10IU／mlDNase Ⅰ獲取神經(jīng)元單細胞懸液，以加入2％B27，5％胎牛血清，1％Glutamax的Neurobasal Medium作為接種及維持培養(yǎng)基，接種48小時加入阿糖胞苷抑制膠質(zhì)細胞生長，綜合使用上述方法建立體外神經(jīng)元培養(yǎng)體系； 2．使用Tubulin β Ⅲ單克隆抗體標記神經(jīng)元，GFAP多克隆抗體標記星型膠質(zhì)細胞，運用免疫熒光技術(shù)檢測胚鼠腦皮質(zhì)神經(jīng)元純度；3．使用國產(chǎn)厭氧培養(yǎng)箱充入85％N2、5％CO2、10％H2混合氣建立胚鼠腦皮質(zhì)神經(jīng)元的缺氧模型；4．倒置相差顯微鏡下觀察缺氧不同時點胚鼠腦皮質(zhì)神經(jīng)元，，對神經(jīng)元缺氧模型進行形態(tài)學評價。
[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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