Name: screening assay for oxidative stress in a feline astrocyte cell line, g355-5
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1。セルの猫アストロサイトの培養およびH 2 O 2による治療<ol><li> 37 DMEMのメディアに加え、栄養素が75 cm 2のフラスコでの培養細胞株G355 - 5℃、5％CO 2 60％コンフルエント（〜5日）まで。すべての1〜2日メディアを交換するかなど、健全な文化を維持するために必要。 </li><li>メディアを取り出して、0.25％トリプシン液2 mlを加える。 45秒のためにピペッティング...

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神経損傷を誘発したウイルスの多くの場合、提案されたメカニズムは、酸化ストレス1、4、7、9、13、15、16、18-22、27、31です。基本的に、それが神経細胞に有毒であるなどの水酸化物、スーパーオキシドアニオン、一酸化窒素、および/または過酸化水素、などのウイルス暴露によるグリア細胞（アストロサイトとミクログリア）放出、活性酸素ラジカル、ことが提案されている。同様...

<table border="1"><tbody><tr><th>試薬の名前</th><th>会社</th><th>カタログ番号</th><th>コメント（省略可能） </th></tr><tr><td> DMEM </td><td> Cellgro </td><td> 15から013 - CV </td><td></td></tr><tr><td> MEMビタミン</td><td> Cellgro </td><td> 25から020 - CL </td><td> 1％（v / v）の</td></tr><tr><td> L -グルタミン</td><td> Hyclone </td><td> 17 - 605E </td><td> 1％（v / v）の</td></tr><tr><td>非必須アミノ酸</td><td> Hyclone </td><td> 25から025 - CL </td><td> 1％（v / v）の</td></tr><tr><td> FBS </td><td> Hyclone </td><td> SH30071.03 </td><td> 20％（v / v）の</td></tr><tr><td>必須アミノ酸</td><td> Cellgro </td><td> 25から030 - CL </td><td> 0.2％（v / v）の</td></tr><tr><td> 500ミリリットル真空ろ過システム</td><td> VWR </td><td> 87006-076 </td><td></td></tr><tr><td> 15 mlコニカルチューブ</td><td>ファルコン</td><td> 352097 </td><td></td></tr><tr><td> 75センチメートル2組織培養フラスコ</td><td>ファルコン</td><td> 353136 </td><td></td></tr><tr><td> 6ウェル組織培養プレート</td><td>ファルコン</td><td> 353224 </td><td></td></tr><tr><td> CM - H 2 DCFDA </td><td>インビトロジェン</td><td> C6827 </td><td></td></tr><tr><td>ヨウ化プロピジウム（PI） </td><td>シグマ</td><td> P4170 - 10mgの</td><td></td></tr><tr><td>トリプシン</td><td>ロンザ</td><td> 17 - 160F </td><td></td></tr><tr><td> H 2 O 2 </td><td>シグマ</td><td> H6520 </td><td></td></tr><tr><td>抗生物質HYQ </td><td> Hyclone </td><td> SV30079.01 </td><td> 0.1％（v / v）の</td></tr><tr><td> G355 - 5細胞</td><td> ATCC </td><td> CRL - 2033 </td><td>通常の猫の脳</td></tr></tbody></table>

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

神経損傷を誘発したウイルスの多くの場合、提案されたメカニズムは、酸化ストレスです。星状膠細胞は、中枢神経系（CNS）の酸化ストレスの制御に重要な役割を担っている。星状膠細胞は、活性酸素種（ROS）から神経細胞を保護するだけでなく、神経細胞のための恒常的な環境を維持するのに役立ちます。 CM - H2DCFDAは、ROSの存在のための細胞透過性の指標です。 CM - H 2 DCFDAは非蛍光化合物として細胞に入り、そして細胞のエステラーゼが酢酸基を除去した後、蛍光灯になり、そして化合物が酸化される。緑色の蛍光であることが判明しているフローサイトメトリーによって測定された細胞の数は、、酸化状態にある細胞の数を示しています。 CM - H 2 DCFDAは異なるROSの多数による酸化を受けやすくなります。このアッセイは関係なく、酸素の酸化的細胞環境のために画面に使用できるようにROSがCM - H 2 DCFDAを酸化させることができるかに関する特異性の欠如は、、、この化合物病因調査の初期段階で使用するための貴重な摂政ですラジカルやROSの組み合わせは、細胞の条件のための責任があります。それは、ROSがCM - H 2 DCFDAの酸化によって存在することが確立されると、その後の追加実験がROSやロスの組み合わせが特定の病因のプロセスに関係しているかを判定するために実施することができます。この研究の結果は、過酸化水素の添加によりCM - H 2 DCFDAの増加はこのアッセイは、G355 - 5細胞内酸化的環境を検出するための貴重なテストであることを示す、生理食塩水対照と比較して検出された蛍光を発することを示しているネコのアストロサイト細胞株。

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

酸化的細胞の環境を検出するスクリーニング法は、CM - H2DCFDAの酸化を測定することです。かつて非蛍光からの蛍光化合物に細胞、CM - H2DCFDA変化の中で酸化する。蛍光の変化はフローサイトメトリーで測定し、酸化的環境における細胞の数を示しています。

猫のアストロサイト細胞株における酸化ストレスのためのスクリーニングアッセイ、G355 - 5

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

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Neuroscience

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

リアルタイム神経細胞株に神経毒性および神経保護に関与する分子経路を描写するツールとしてのインピーダンスベースの細胞分析装置

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

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

間接ニューロン、星状細胞共培養アッセイ：<em&gt;インビトロ</em&gt;ニューロングリアの相互作用の詳細な調査のためのセットアップ

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

従来のUVレーザー共焦点顕微鏡を用いて細胞ストレスと死のトリガ

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

ビオチン化を用いた初代星状細胞培養におけるタンパク質の細胞表面発現およびエンドサイトーシス速度の測定

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

Sod1 突然変異体蛋白質集計における細胞の高いコンテンツ定量化のための分析法の開発

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

貪食能アッセイのアストロ サイトを介した新規体外ライブ イメージングを使用して pH インジケーター共役シナプトソーム

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

体外タウ蛋白と創薬スクリーニングの集計を勉強するための試金

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

ゼブラフィッシュ稚魚における側線有毛細胞の生体内カルシウム イメージング

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

ネコ大型動物モデルにおけるヒト胚性幹細胞由来網膜組織の網膜下移植

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

変異型メチオニンtRNA合成酵素マウス株を用いた複合体組織からの細胞型特異的タンパク質精製・同定

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