本文選題：阿糖胞苷　切入點(diǎn)：大腦皮質(zhì)　出處：《中國組織工程研究》2017年12期

【摘要】：背景:由于中樞神經(jīng)系統(tǒng)疾病研究需求,實(shí)驗(yàn)室常用有毒性的阿糖胞苷獲得純度較高的神經(jīng)元細(xì)胞,然而阿糖胞苷的介入時(shí)間鮮見研究。目的:驗(yàn)證終濃度為10μmol/L阿糖胞苷在大鼠皮質(zhì)神經(jīng)元原代細(xì)胞培養(yǎng)中的適宜介入時(shí)間。方法:采用神經(jīng)元專用培養(yǎng)基Neurobasal+B27進(jìn)行大腦皮質(zhì)組織原代神經(jīng)元培養(yǎng),分別于接種細(xì)胞后12,24,36,48 h加入終濃度為10μmol/L阿糖胞苷。每48 h半量換液。于7 d后倒置的顯微鏡下觀察神經(jīng)元形態(tài);采用免疫細(xì)胞化學(xué)的方法對(duì)神經(jīng)元特異性烯醇化酶進(jìn)行免疫化學(xué)染色,鑒定神經(jīng)元細(xì)胞的純度和成熟度;計(jì)算機(jī)多功能圖像分析系統(tǒng)對(duì)所有神經(jīng)元特異性烯醇化酶陽性細(xì)胞進(jìn)行形態(tài)學(xué)計(jì)量分析。結(jié)果與結(jié)論:(1)接種細(xì)胞后24 h加入阿糖胞苷,神經(jīng)元純度在90%以上,分化成熟神經(jīng)元細(xì)胞占比89.00%,神經(jīng)元細(xì)胞胞體面積最大,突觸最長,胞質(zhì)透亮,核大而明顯,細(xì)胞體周圍折光性強(qiáng),立體感強(qiáng),突起三四個(gè),具有三四級(jí)分叉,形成良好的網(wǎng)絡(luò)結(jié)構(gòu),非神經(jīng)元細(xì)胞較少,神經(jīng)元純度及細(xì)胞狀態(tài)最佳;(2)結(jié)果表明,阿糖胞苷介入培養(yǎng)過程越早,神經(jīng)元細(xì)胞純度越高,但阿糖胞苷對(duì)神經(jīng)元細(xì)胞分化成熟的影響也會(huì)越明顯;采用neurolbasal培養(yǎng)基,于接種細(xì)胞后24 h加入阿糖胞苷,可以獲得理想純度,成熟度較好,生長狀態(tài)較好的大腦皮質(zhì)神經(jīng)元。
[Abstract]:Background: due to the need of central nervous system disease research, the toxic cytarabine is commonly used in laboratory to obtain high purity neuronal cells. However, the interventional time of cytarabine is rarely studied.Aim: to investigate the optimal interventional time of cytarabine (10 渭 mol/L) in primary culture of rat cortical neurons.Methods: primary neurons of cerebral cortex were cultured on Neurobasal B27 neuron culture medium. The final concentration of cytarabine was 10 渭 mol/L after inoculation.The liquid was changed every 48 hours.The morphology of neurons was observed under inverted microscope 7 days later and the purity and maturity of neuron cells were identified by immunocytochemical staining of neuron-specific enolase.The morphometric analysis of all neuron-specific enolase positive cells was performed by computer multifunctional image analysis system.Results and conclusion 24 hours after inoculation, cytarabine was added to the cells, the purity of neurons was over 90%, the proportion of differentiated neurons was 89.00, the area of neuronal cell body was the largest, the synapse was longest, the cytoplasm was clear, the nucleus was large and obvious.The results showed that there were three or four processes with three or four degrees of bifurcation, a good network structure, fewer non-neuron cells, and the best purity and state of the neurons.The earlier the cytarabine interventional culture process, the higher the purity of neuronal cells, but the more obvious the effect of cytarabine on the differentiation and maturation of neuronal cells. Cytarabine was added in neurolbasal medium 24 hours after inoculation.The cortical neurons with ideal purity, good maturity and good growth state can be obtained.
【作者單位】： 保定市第一中心醫(yī)院神經(jīng)外科;河北大學(xué)醫(yī)學(xué)院;北京理工大學(xué)生命學(xué)院;解放軍軍事交通學(xué)院第九隊(duì);
【基金】：河北省自然科學(xué)基金(B2015201161) 國家自然科學(xué)基金(81502477)~~
【分類號(hào)】：R741
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本文編號(hào)：1708868