Name: cellular toxicity of nanogenomedicine in mcf-7 cell line: mtt assay
Uploaded: 2009-04-03T00:00:00
Description: Watch this Scientific Journal Video about Cellular Toxicity of Nanogenomedicine in MCF-7 Cell Line: MTT assay at JoVE.com

<p class="jove_content">著者らは、贈り物として注射用フェノールレッドおよび滅菌水を含まないDMEMを提供するため、それぞれの幹細胞技術とシャヒガジタブリーズの企業に感謝します。 </p>

<p class="jove_title"> MCF - 7を96ウェルプレートに播種。 </p><p class="jove_content"> MCF - 7細胞は、37ダルベッコ変法イーグル培地（DMEM）、10％FBS、100 U / mlペニシリン、および100μg/ mlのストレプトマイシンを含む培地中で25 T -フラスコで培養し、℃、5％CO <sub>2、95％</sub>空気と完全な湿度。一度〜90％コンフルエントに達した、彼らを0.05％トリプシン/ EDTAを用いて分離し、トリパンブルーや血球計算盤により計数した。こ...

<ol>
	<li>Plumb, J. A., Milroy, R., Kaye, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+the+pH+Dependence+of+3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium+Bromide-Formazan+Absorption+on+Chemosensitivity+Determined+by+a+Novel+Tetrazolium-based+Assay.">Effects of the pH Dependence of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide-Formazan Absorption on Chemosensitivity Determined by a Novel Tetrazolium-based Assay.</a> <em style='font-style: italic'>Cancer Research</em>. <b>49</b>, 4435-4435 (1989).</li><li>Mosmann, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+Colorimetric+Assay+for+Cellular+Growth+and+Survival:+Application+to+Proliferation+and+Cytotoxicity+Assays.">Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assays.</a> <em style='font-style: italic'>Journal of Lmmunological Methods</em>. <b>65</b>, 55-55 (1983).</li><li>Yang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Energy+and+Redox+States+in+the+C6+Glioma+Cells+Following+Acute+Exposure+to+Zn,+Se++4+,+and+Se++6+and+the+Correlation+with+Apoptosis.">Energy and Redox States in the C6 Glioma Cells Following Acute Exposure to Zn, Se +4 , and Se +6 and the Correlation with Apoptosis.</a> <em style='font-style: italic'>Toxicology Mechanisms and Methods</em>. <b>16</b> (1), 13-13 (2006).</li><li>Spinner, D. M., Bartlett, J. M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>MTT Growth Assays in Ovarian Cancer, in Ovarian Cancer: Methods and Protocols</em>. , 175-177 (2000).</li></ol>

<p class="jove_content"> MTTアッセイは、汎用的な方法とみなされ、それに応じて細胞の生存率は、様々な治療法により評価することができた。結果ホルマザンの産生は細胞内のエネルギー代謝のレベルに比例するように見えます。したがって、それも細胞増殖がない場合に代謝的に活性化された細胞を測定することが可能です。生成されたホルマザンの量は、培養培地中のMTTの量に比例する。一方で、生産ホルマザ...

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		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<td>25 t-flask</td>
<td>&nbsp;</td>
<td>Orange Scientific</td>
<td>5510100</td>
<td>&nbsp;</td>
<tr>
<td>PBS</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>P3813</td>
<td>&nbsp;</td>
<tr>
<td>Dulbecco&#x2019;s Modified Eagle&#x2019;s Medium - high glucose</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>D5671</td>
<td>&nbsp;</td>
<tr>
<td>MTT</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>M2128</td>
<td>&nbsp;</td>
<tr>
<td>FBS</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>F2442</td>
<td>&nbsp;</td>
<tr>
<td>Penicillin 200,000 u/ml</td>
<td>&nbsp;</td>
<td>CinnaGen</td>
<td>CR7913</td>
<td>&nbsp;</td>
<tr>
<td>Streptomycin 200 mg/ml</td>
<td>&nbsp;</td>
<td>CinnaGen</td>
<td>CR7912</td>
<td>&nbsp;</td>
<tr>
<td>trypsin/EDTA</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>T3924</td>
<td>&nbsp;</td>
<tr>
<td>trypan blue</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>T8154</td>
<td>&nbsp;</td>
<tr>
<td>Superfect</td>
<td>&nbsp;</td>
<td>Qiagen</td>
<td>301105</td>
<td>&nbsp;</td>
<tr>
<td>Polyfect</td>
<td>&nbsp;</td>
<td>Qiagen</td>
<td>301305</td>
<td>&nbsp;</td>
<tr>
<td>EGFR antisense</td>
<td>&nbsp;</td>
<td>MWG biotech</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Scrambled antisense</td>
<td>&nbsp;</td>
<td>MWG biotech</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>DMEM without phenol red</td>
<td>&nbsp;</td>
<td>Gibco</td>
<td>11880</td>
<td>&nbsp;</td>
<tr>
<td>Glycine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>G8898</td>
<td>&nbsp;</td>
<tr>
<td>hemocytometer</td>
<td>&nbsp;</td>
<td>HBG </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>8-channel pipette</td>
<td>&nbsp;</td>
<td>TreffLab Transferpette</td>
<td>96.9918.10.1</td>
<td>&nbsp;</td>
<tr>
<td>Disposable syringe filter </td>
<td>&nbsp;</td>
<td>IWAKI</td>
<td>2052-025</td>
<td>&nbsp;</td>
<tr>
<td>Plate reader equipped with 570 nm &630 nm filters </td>
<td>&nbsp;</td>
<td>Biotek ELx808</td>
<td>ELX808</td>
<td>&nbsp;</td>
<tr>
<td>96-well plate with lid (flat bottom)</td>
<td>&nbsp;</td>
<td>IWAKI</td>
<td>3860-096</td>
<td>&nbsp;</td>
<tr>
<td>Laminar flow hood</td>
<td>&nbsp;</td>
<td>Esco class II Biosafety Cabinet</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>CO<sub>2</sub> Incubator</td>
<td>&nbsp;</td>
<td>ASTEC</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
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cellular toxicity of nanogenomedicine in mcf-7 cell line: mtt assay

<p class="jove_content">未来nanogenomedicine（例えば、短干渉RNAおよびアンチセンス）の細胞毒性は、臨床開発を妨げる可能性があります。これらのうち、遺伝子ベースの医学および/またはそのキャリアは、細胞毒性を誘発し得る。このような細胞毒性の評価のために、一般的な方法論は、3の利用率に大きく依存します - として広く使用されている（4,5 - ジメチル-2 - チアゾリル）-2、5 - ジフェニル- 2H -テトラゾリウムブロマイド（MTT）アッセイ細胞内のミトコンドリアのデヒドロゲナーゼの酵素の活性に基づいて比色分析的なアプローチ。この現在の調査では、MCF - 7細胞を96ウェルプレートにし、50％コンフルエントで接種された彼らは、24時間後に別のnanopolyplexesで処理し、MTTアッセイを行った。水溶性黄色のMTTは、さらにジメチルスルホキシドとSornsonの緩衝液pH 10.5に溶解して水不溶性の紫色ホルマザンへの代謝活性を有する細胞で代謝される。得られた生成物は、570nmでプレートリーダーを用いて分光光度法により定量することができる。</p>

Watch this Scientific Journal Video about Cellular Toxicity of Nanogenomedicine in MCF-7 Cell Line: MTT assay at JoVE.com

Cellular Toxicity of Nanogenomedicine in MCF-7 Cell Line: MTT assay

<p class="jove_content">MTTアッセイは、水不溶性の紫色ホルマザンの黄色のMTTと生産の減少に基づいて細胞生存率の評価のための簡単​​かつ再現可能な比色アッセイである。ここでは、nanogenomedicineの治療時にMCF - 7細胞の生存率が評価されています。</p>

MCF - 7細胞株におけるNanogenomedicineの細胞毒性：MTTアッセイ

Cytotoxicity of the futuristic nanogenomedicine (e.g., short interfering RNA and antisense) may hamper its clinical development. Of these, the gene-based ...

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