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"> 저자는 선물로 주입에 대한 페놀 빨간색과 멸균 물없이 DMEM을 제공하는 각각 줄기 세포 기술과 샤히드 Ghazi 타브 리즈 회사를 인정하고 싶습니다. </p>

<p class="jove_title"> 96 - 웰 플레이트에 MCF - 7 시딩 : </p><p class="jove_content"> MCF - 7 세포는 37 Dulbecco의 수정된 이글의 중간 (DMEM) 10 % FBS, 100 U / ML 페니실린, 100 μg / ML 스트렙토 마이신을 포함하는 배지에서 25 T - 플라스크에 교양되었습니다 ° C 5% CO <sub>2,</sub> 95 %를 공기와 습기 완료. 일단 ~ 90 % confluency에 도달, 그들은 0.05 % 트립신 / EDTA (에틸렌 다이아 민 테트라 초산)를 사용하여 분리하고 trypan 파란색과 hemocytometer에 의해 ...

<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 분석은 다양한 방법으로 간주하고 이에 따라 세포의 생존은 다양한 치료시 평가 수 있습니다. 결과 formazan의 생산은 세포의 에너지 대사의 수준에 비례하는 것 같아요. 따라서도 세포 증식의 부재에 metabolically 활성 세포를 측정할 수 있습니다. 생산 formazan의 양은 배양 매체에서 MTT의 금액에 비례합니다. 동안 formazan의 최대 금액을 달성하는 데 필요한 MTT의 농도는 다른 세포 라인의 활용도에 따...

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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>
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<td>Dulbecco&#x2019;s Modified Eagle&#x2019;s Medium - high glucose</td>
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<td>D5671</td>
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<td>MTT</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>M2128</td>
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<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>F2442</td>
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<td>Sigma-Aldrich</td>
<td>T8154</td>
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<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>
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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>
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<td>Gibco</td>
<td>11880</td>
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<td>HBG </td>
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<td>8-channel pipette</td>
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<td>TreffLab Transferpette</td>
<td>96.9918.10.1</td>
<td>&nbsp;</td>
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<td>Disposable syringe filter </td>
<td>&nbsp;</td>
<td>IWAKI</td>
<td>2052-025</td>
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<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>
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<tr>
<td>Laminar flow hood</td>
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<td>Esco class II Biosafety Cabinet</td>
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<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">미래 nanogenomedicine (예, 짧은 간섭 RNA와 안티 센스)의 세포 독성은 임상 개발을 방해 수 있습니다. 이들 중, 유전자 기반의 의학 및 / 또는 이동 통신사에서 세포 독성을 이끌어 수 있습니다. 이러한 세포 독성 평가를위한 일반적인 방법론 3의 활용에 따라 크게 좌우된다 - (4, 5 - 디메틸 - 2 - thiazolyl) 널리 사용되었습니다 -2, 5 - 디페닐 - 2 시간 - tetrazolium 브로마이드 (MTT) 분석 세포 mitochondrial 탈수소 효소의 활동을 기반으로하는 colorimetric 접근. 이 현재 조사에서, MCF - 7 세포는 96 - 웰 플레이트에 주사했다 50 % confluency에서 그들은 서로 다른 nanopolyplexes 대우했다, 24 시간 후 MTT 분석을 들어서는. 수용성 황색 MTT는 더욱 dimethylsulfoxide 및 Sornson의 버퍼 산도 10.5에 녹아있는 물에 불용성 보라색 formazan에 metabolically 활성 세포에 의해 대사됩니다. 결과 제품은 570 nm의에서 플레이트 판독기를 사용하여 분광 광도법으로 계량하실 수 있습니다.</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 분석은 노란색 MTT의 감소와 물 불용성 보라색 formazan의 생산을 기반으로 세포 생존 능력 평가를위한 간단하고 재현성 colorimetric 분석이다. 여기 nanogenomedicine 치료시 MCF - 7 세포의 생존 능력이 평가되었습니다.</p>

MCF - 7 세포주에서 세포 독성 Nanogenomedicine의 : MTT 분석

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