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">作者想承认干细胞技术和大不里士沙希德亚瓦尔公司分别提供DMEM培养液无酚红和无菌水注射作为礼物。 </p>

<p class="jove_title"> MCF - 7播种在96孔板： </p><p class="jove_content"> MCF - 7细胞分别培养在25吨的烧瓶中包含在37贝科的修改鹰的中等（DMEM），10％胎牛血清，100单位/毫升青霉素，和100微克/毫升链霉素中等°彗星与5％的CO <sub>2，95％</sub>空气和完整的湿度。一旦达到〜90％汇合，他们分离用0.05％胰蛋白酶/ EDTA，并通过台盼蓝和血球计数。这些细胞，然后悬浮在浓度为4 × 10 <sup>4</sup>细胞/厘米<sup>2，</sup>补充到96孔板，8道移?...

<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法被认为是一个通用的方法，可以根据各种治疗方法的评估和相应的细胞的可行性。由此产生的甲臜的生产似乎是在细胞能量代谢水平成正比。因此，它是可能的措施，甚至在细胞增殖的情况下的代谢活性的细胞。产生的甲臜量在孵化介质的MTT量是成正比的。虽然，这是要求达到最高金额的甲臜的MTT浓度产生的可能发生变化后，利用不同的细胞系。此外，使用这种检测，可以检测到非常小的活...

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		<th>Catalogue Number</th>
		<th>Comment</th>
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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>
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cellular toxicity of nanogenomedicine in mcf-7 cell line: mtt assay

<p class="jove_content">未来派nanogenomedicine（例如，短干扰RNA和反义）的细胞毒性，可能妨碍其临床发展。这些基因为基础的医药和/或它的载体可能会引起细胞毒性。对于评估等的细胞毒作用，常见的方法是在很大程度上依赖利用3 - （4,5 - 二甲基-2 - 噻唑基）-2，5 - 二苯基- 2H -四氮唑溴（MTT）法已被广泛用作比色的方法，在细胞线粒体脱氢酶的酶的活性的基础上。在当前的调查，MCF - 7细胞接种在96孔板和50％汇合，他们与不同的nanopolyplexes对待，并受到24小时后，以MTT法检测。水溶性的黄色的MTT代谢的代谢活跃的细胞，不溶于水的紫色的甲臜，这是进一步溶解在二甲基亚砜和Sornson的缓冲区pH值10.5。由此产生的产品是可以量化的使用酶标仪在570 nm处一个分光。</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法是一种简便，重现性好比色法测定细胞活力评价的基础上减少黄色的MTT和不溶于水，紫色的甲臜的生产。在这里，nanogenomedicine治疗后，MCF - 7细胞的可行性进行了评估。</p>

Nanogenomedicine在MCF - 7细胞的细胞毒性：MTT法

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