Name: screening assay for oxidative stress in a feline astrocyte cell line, g355-5
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1。猫星形胶质细胞的细胞培养和治疗与H 2 O 2 <ol><li>文化细胞系G355 - 5在75厘米的 2瓶用DMEM媒体加营养素在37 ° C和5％的CO 2，直到60％融合（〜5天）。每1-2天更换介质或需要保持一个健康的文化。 </li><li>取出介质，加2毫升0.25％胰蛋白酶。电梯45秒吹打细胞 - 在室温（RT）1分钟。 </li><li>快速传输悬挂一个15毫升的锥形管中，含有10毫升的媒体。轻轻混匀。 </li><li>离心3分钟?...

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通常建议的病毒的机制诱导神经元的损伤是氧化应激1，4，7，9，13，15，16，18-22，27，31 。基本上，它是通过病毒暴露的神经胶质细胞（星形胶质细胞和小胶质细胞）释放，如氧化钾，超氧阴离子自由基，一氧化氮，和/或过氧化氢，这是有毒的神经细胞的活性氧自由基。同样，氧化应激也被作为一个主要的甲基苯丙胺神经毒性2，6，17的病理机制的建议。由于本集团在病毒和药物?...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> DMEM培养液</td><td> Cellgro </td><td> 15 - 013 - CV </td><td></td></tr><tr><td>纪念维生素</td><td> Cellgro </td><td> 25 - 020 - CL </td><td> 1％（V / V） </td></tr><tr><td> L -谷氨酰胺</td><td> Hyclone公司</td><td> 17 - 605E </td><td> 1％（V / V） </td></tr><tr><td>非必需氨基酸</td><td> Hyclone公司</td><td> 25 - 025 - CL </td><td> 1％（V / V） </td></tr><tr><td>胎牛血清</td><td> Hyclone公司</td><td> SH30071.03 </td><td> 20％（V / V） </td></tr><tr><td>人体必需的氨基酸</td><td> Cellgro </td><td> 25 - 030 - CL </td><td> 0.2％（V / V） </td></tr><tr><td> 500毫升真空过滤系统</td><td> VWR </td><td> 87006-076 </td><td></td></tr><tr><td> 15 ml锥形管</td><td>鹘</td><td> 352097 </td><td></td></tr><tr><td> 75厘米2组织培养瓶</td><td>鹘</td><td> 353136 </td><td></td></tr><tr><td> 6孔细胞培养板</td><td>鹘</td><td> 353224 </td><td></td></tr><tr><td> CM - H 2 DCFDA </td><td> Invitrogen公司</td><td> C6827 </td><td></td></tr><tr><td>碘化丙啶（PI） </td><td>西格玛</td><td> P4170 - 10毫克</td><td></td></tr><tr><td>胰蛋白酶</td><td>龙沙</td><td> 17 - 160F </td><td></td></tr><tr><td> H 2 O 2 </td><td>西格玛</td><td> H6520 </td><td></td></tr><tr><td> HYQ抗生素</td><td> Hyclone公司</td><td> SV30079.01 </td><td> 0.1％（V / V） </td></tr><tr><td> G355 - 5细胞</td><td> ATCC </td><td> CRL - 2033 </td><td>正常猫科动物的大脑</td></tr></tbody></table>

screening assay for oxidative stress in a feline astrocyte cell line, g355-5

的病毒通常建议的机制，诱导神经元的损伤是氧化应激。星形胶质细胞有重要作用，在控制对中枢神经系统（CNS）的氧化应激。星形胶质细胞有助于保持神经元以及保护神经元的活性氧（ROS）的稳态环境。 CM - H2DCFDA是一个细胞的活性氧的存在透水的指标。 CM - H 2 DCFDA作为一个非荧光化合物进入细胞，并成为荧光细胞酯酶删除后的醋酸组，该化合物是氧化。细胞，流式细胞仪检测，发现绿色荧光细胞处于氧化状态的指示。 CM - H 2 DCFDA是由不同的ROS的大量氧化易感。这种缺乏特异性，对活性氧可以氧化CM - H 2 DCFDA，使该化合物为使用宝贵的摄政在一个发病调查的早期阶段，这种检测可以用于氧化的细胞环境到屏幕上，无论是其中的氧气自由基或活性氧相结合是负责细胞的条件。一旦它被建立，活性氧是目前CM - H 2 DCFDA氧化，然后可以进行更多的实验来确定在特定的发病过程中所涉及ROS或组合罗斯。这项研究的结果表明，与过氧化氢 ​​增加CM - H 2 DCFDA的加入荧光检测与生理盐水对照，表明这个实验是一个宝贵的测试，检测G355 - 5细胞内的氧化环境猫星形胶质细胞株。

Watch this Scientific Journal Video about Screening Assay for Oxidative Stress in a Feline Astrocyte Cell Line, G355-5 at JoVE.com

Screening Assay for Oxidative Stress in a Feline Astrocyte Cell Line, G355-5

一种检查方法检测细胞氧化的环境，来衡量的CM - H2DCFDA氧化。一旦氧化细胞内，CM - H2DCFDA的变化，从非荧光一​​种荧光的化合物。是用流式细胞仪检测荧光的这种变化，并表示在氧化环境中的细胞数量。

筛选法测定氧化应激在猫的星形胶质细胞的细胞线，G355 - 5

An often-suggested mechanism of virus induced neuronal damage is oxidative stress. Astrocytes have an important role in controlling oxidative stress of ...

Research

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Real-Time Impedance-based Cell Analyzer as a Tool to Delineate Molecular Pathways Involved in Neurotoxicity and Neuroprotection in a Neuronal Cell Line

作为一种工具来描绘分子途径参与神经毒性和神经保护神经元细胞在线实时阻抗为基础的细胞分析仪

Many brain-related disorders have neuronal cell death involved in their pathophysiology. Improved in vitro models to study neuroprotective or neurotoxic ...

科研

神经科学

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

间接神经元的星形胶质细胞共培养试验：一个<em&gt;体外</em&gt;设置 - 为神经元的神经胶质细胞相互作用的详细调查

Proper neuronal development and function is the prerequisite of the developing and the adult brain. However, the mechanisms underlying the highly ...

Triggering Cell Stress and Death Using Conventional UV Laser Confocal Microscopy

触发细胞应激和死亡使用传统的紫外线激光共聚焦显微镜

Using a standard confocal setup, a UV ablation method can be utilized to selectively induce cellular injury and to visualize single-cell responses and ...

Determining Cell-surface Expression and Endocytic Rate of Proteins in Primary Astrocyte Cultures Using Biotinylation

使用生物素化测定原代星形细胞培养物中蛋白质的细胞表面表达和吞噬率

Cell-surface proteins mediate a wide array of functions. In many cases, their activity is regulated by endocytic processes that modulate their levels at ...

Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells

活细胞 Sod1 突变蛋白聚集物高含量定量测定方法的建立

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that can be caused by inherited mutations in the gene encoding copper-zinc ...

A Novel In Vitro Live-imaging Assay of Astrocyte-mediated Phagocytosis Using pH Indicator-conjugated Synaptosomes

A Novel In Vitro Live-imaging Assay of Astrocyte-mediated Phagocytosis Using pH Indicator-conjugated Synaptosomes

一种新的体外活体成像法测定星形胶质细胞的吞噬作用 pH 指标-共轭突

Astrocytes are the major cell type in the brain and directly contact synapses and blood vessels. Although microglial cells have been considered the major ...

In Vitro Assay for Studying the Aggregation of Tau Protein and Drug Screening

In Vitro Assay for Studying the Aggregation of Tau Protein and Drug Screening

在体外陶氏蛋白聚集与药物筛选研究方法

Aggregation of tau protein and formation of paired helical filaments is a hallmark of Alzheimer&#39;s disease and other tauopathies. Compared to other ...

In Vivo Calcium Imaging of Lateral-line Hair Cells in Larval Zebrafish

幼虫斑马鱼外系头发细胞的活体钙成像

Sensory hair cells are mechanoreceptors found in the inner ear that are required for hearing and balance. Hair cells are activated in response to sensory ...

Subretinal Transplantation of Human Embryonic Stem Cell-Derived Retinal Tissue in a Feline Large Animal Model

猫大型动物模型中人类胚胎干细胞来源的视网膜组织的视网膜下移植

Retinal degenerative (RD) conditions associated with photoreceptor loss such as age-related macular degeneration (AMD), retinitis pigmentosa (RP) and ...

Cell-Type Specific Protein Purification and Identification from Complex Tissues Using a Mutant Methionine tRNA Synthetase Mouse Line

Understanding protein homeostasis in vivo is key to knowing how the cells work in both physiological and disease conditions. The present protocol ...