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"> Yazarlar, hediye olarak enjeksiyon için fenol kırmızısı ve steril su olmadan DMEM sağlamak için sırasıyla Kök Hücre Teknolojisi ve Shahid Gazi Tebriz şirketlerinin kabul etmek istiyorum. </p>

<p class="jove_title"> MCF-7 96-plaka tohumlama: </p><p class="jove_content"> MCF-7 hücreleri 37 ° Dulbecco&#39;nun Modifiye Kartal Kullanıcı Orta (DMEM),% 10 FBS, 100 U / ml penisilin, ve 100 mg / ml streptomisin içeren orta 25 t-balon kültüre edildi ° C&#39;de% 5 CO <sub>2,%</sub> 95 ile hava ve tam nem. Bir kez ~% 90 confluency ulaştı, 0.05% tripsin / EDTA ile müstakil ve tripan mavi ve hemasitometre sayılır. Bu hücreler daha sonra 4 × 10 <sup>4</sup> hücre / cm <sup>2</sup> &#39;lik bir konsantrasyon yeniden bekletildi ve 8 ...

<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 testi, çok yönlü bir yöntem olarak kabul edilir ve buna göre çeşitli tedaviler üzerine hücrelerin canlılığı değerlendirilebilir. Sonuç formazan üretimi, hücrelerde enerji metabolizması düzeyi ile orantılı olarak görünür. Bu nedenle, metabolik olarak aktif hücrelerin hücre proliferasyonu yokluğunda bile ölçmek mümkündür. Üretilen formazan miktarı inkübasyon orta MTT miktarı ile doğru orantılıdır. Ederken, formazan maksimum miktarını elde etmek için gerekli olan MTT konsantra...

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<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>
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<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>
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<td>Penicillin 200,000 u/ml</td>
<td>&nbsp;</td>
<td>CinnaGen</td>
<td>CR7913</td>
<td>&nbsp;</td>
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<td>Streptomycin 200 mg/ml</td>
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<td>CinnaGen</td>
<td>CR7912</td>
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<td>trypsin/EDTA</td>
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<td>Sigma-Aldrich</td>
<td>T3924</td>
<td>&nbsp;</td>
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<td>trypan blue</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>T8154</td>
<td>&nbsp;</td>
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<td>Superfect</td>
<td>&nbsp;</td>
<td>Qiagen</td>
<td>301105</td>
<td>&nbsp;</td>
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<td>Polyfect</td>
<td>&nbsp;</td>
<td>Qiagen</td>
<td>301305</td>
<td>&nbsp;</td>
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<td>EGFR antisense</td>
<td>&nbsp;</td>
<td>MWG biotech</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Scrambled antisense</td>
<td>&nbsp;</td>
<td>MWG biotech</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>DMEM without phenol red</td>
<td>&nbsp;</td>
<td>Gibco</td>
<td>11880</td>
<td>&nbsp;</td>
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<td>Glycine</td>
<td>&nbsp;</td>
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<td>G8898</td>
<td>&nbsp;</td>
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<td>HBG </td>
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<td>&nbsp;</td>
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<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>
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cellular toxicity of nanogenomedicine in mcf-7 cell line: mtt assay

<p class="jove_content">Fütüristik nanogenomedicine (örneğin, kısa müdahale RNA ve antisens) Sitotoksisite klinik geliştirme engel olabilir. Bunlardan, gen bazlı ilaç ve / veya taşıyıcı hücresel toksisite ortaya çıkarmak. Gibi sitotoksisite değerlendirilmesi için ortak bir metodoloji, 3 kullanımı üzerine büyük ölçüde bağlıdır (4, 5-dimetil-2-thiazolyl) -2, yaygın olarak kullanılan 5-difenil-2H-tetrazolium bromide (MTT) tayini hücrelerinde mitokondriyal dehidrogenaz enzimlerin aktivitesi üzerine dayalı bir kolorimetrik yaklaşım. Bu araştırmada, MCF-7 hücreleri 96-plaka ekildi ve 50% confluency farklı nanopolyplexes ile tedavi edildi ve 24 saat sonra tahlil MTT için tabi. Suda çözünen sarı MTT dimethylsülfoksit ve Sornson s tampon pH 10,5 daha fazla çözünmüş suda çözünmeyen mor formazan, metabolik olarak aktif hücreler tarafından metabolize edilir. Ortaya çıkan ürün, 570 nm&#39;de bir plaka okuyucu kullanarak spektrofotometri tarafından belirlenebilir.</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 testi sarı MTT azaltılması ve suda çözünmeyen mor formazan üretimine dayalı hücre canlılığının değerlendirilmesi için kolay ve tekrarlanabilir kolorimetrik assay. Burada, nanogenomedicine tedavi üzerine MCF-7 hücre canlılığı değerlendirildi.</p>

MCF-7 Hücre Line Nanogenomedicine Hücresel Toksisite: MTT assay

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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