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"> Los autores desean agradecer a la tecnología de células madre y Shahid empresas Ghazi Tabriz, respectivamente, para proveer de agua DMEM sin rojo fenol y estéril para inyección como regalos. </p>

<p class="jove_title"> MCF-7 de siembra en placa de 96 pocillos: </p><p class="jove_content"> MCF-7 células fueron cultivadas en 25 t-matraz en un medio que contiene Dulbecco Modificado Medio Eagle (DMEM), 10% de SFB, 100 penicilina U / ml, y 100 estreptomicina mg / ml a 37 ° C con 5% <sub>CO2,</sub> 95% aire y la humedad total. Una vez llegado a 90% de confluencia, se separaron con 0,05% de tripsina / EDTA y se cuentan por medio de azul de tripano y un hemocitómetro. Estas células se resuspendieron luego en una concentración de 4 × 10 <s...

<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"> El ensayo de MTT se considera que es un método versátil y, en consecuencia la viabilidad de las células podrían ser evaluados en diversos tratamientos. La producción de formazán resultante parece ser proporcional al nivel de metabolismo de la energía en las células. Por lo tanto, es posible medir las células metabólicamente activa, incluso en ausencia de la proliferación celular. La cantidad de formazán producida es proporcional a la cantidad de MTT en el medio de incubación. Mientras, la concentración de ...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<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>
		</div>
	

cellular toxicity of nanogenomedicine in mcf-7 cell line: mtt assay

<p class="jove_content">Citotoxicidad de los nanogenomedicine futurista (por ejemplo, corto de interferencia de ARN antisentido y) puede obstaculizar su desarrollo clínico. De ellos, la medicina basada en genes y / o su compañía pueden provocar toxicidad celular. Para la evaluación de la citotoxicidad como, una metodología común depende en gran medida la utilización de la 3 - (4, 5-dimetil-2-tiazolil) -2, 5-difenil-2H-tetrazolio (MTT) de ensayo que ha sido ampliamente utilizado como un enfoque colorimétrico basado en la actividad de las enzimas deshidrogenasa mitocondrial de las células. En esta investigación actual, células MCF-7 fueron inoculadas en placa de 96 pocillos y en la confluencia del 50% fueron tratados con nanopolyplexes diferentes y sometidos a ensayo MTT después de 24 horas. Agua amarilla MTT soluble es metabolizado por las células metabólicamente activas para el agua insoluble formazán púrpura, que se ve disuelta en buffer de pH dimetilsulfóxido y Sornson s 10,5. El producto resultante puede ser cuantificado por espectrofotometría utilizando un lector de placas a 570 nm.</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">El ensayo de MTT es un ensayo colorimétrico fácil y reproducible para la evaluación de la viabilidad celular basado en la reducción de MTT amarilla y la producción de agua morada formazán insoluble. En este caso, la viabilidad de las células MCF-7 sobre el tratamiento de nanogenomedicine ha sido evaluada.</p>

La toxicidad celular de Nanogenomedicine en la línea celular MCF-7: ensayo de MTT

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

Research

JoVE Journal

Biology

Proboscis Extension Response (PER) Assay in Drosophila

Proboscis Extension Response PER Assay in Drosophila

Extensión probóscide Respuesta (PER) Ensayo en Drosophila

Proboscis extension response (PER) is a taste behavior assay that has been used in flies as well as in honeybees.
On the surface of the fly's mouth ...

Investigación

Biología

Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)

Multiplex PCR and Reverse Line Blot Hybridization Assay mPCR/RLB

Multiplex PCR y Blot Línea ensayo de hibridación reversa (MPCR / RLB)

Multiplex PCR/Reverse Line Blot Hybridization assay allows the detection of up to 43 molecular targets in 43 samples using one multiplex PCR reaction ...

Long-term Lethal Toxicity Test with the Crustacean <em>Artemia franciscana</em>

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

A largo plazo Prueba de toxicidad letal con el Crustáceo<em> Artemia franciscana</em

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish ...

Measurement of Cellular Chemotaxis with ECIS/Taxis

La medición de la quimiotaxis celular con ECIS / taxis

Cellular movement in response to external stimuli is fundamental to many cellular processes including wound healing, inflammation and the response to ...

Organelle Transport in Cultured <em>Drosophila</em> Cells: S2 Cell Line and Primary Neurons.

Organelle Transport in Cultured Drosophila Cells: S2 Cell Line and Primary Neurons.

Orgánulos Transporte en Cultivadas<em&gt; Drosophila</em&gt; Celdas: S2 Línea Celular y neuronas primarias.

Drosophila S2 cells plated on a coverslip in the presence of any actin-depolymerizing drug form long unbranched processes filled with uniformly polarized ...

An Assay for Lateral Line Regeneration in Adult Zebrafish

Un ensayo para la línea lateral de regeneración en adultos de pez cebra

Due to the clinical importance of hearing and balance disorders in man, model organisms such as the zebrafish have been used to study lateral line ...

One-channel Cell-attached Patch-clamp Recording

Un canal celular conectado a Patch-clamp grabación

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate ...

Generating a &#34;Humanized&#34; <em>Drosophila</em> S2 Cell Line Sensitive to Pharmacological Inhibition of Kinesin-5

Generating a "Humanized" Drosophila S2 Cell Line Sensitive to Pharmacological Inhibition of Kinesin-5

Generación de un &quot;humanizado&quot;<em&gt; Drosophila</em&gt; S2 Línea Celular Sensible a farmacológico Inhibición de la Kinesin-5

Kinetochores are large protein-based structures that assemble on centromeres during cell division and link chromosomes to spindle microtubules. Proper ...

A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain (RTA) Trafficking in Yeast

A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain RTA Trafficking in Yeast

Un análisis Simple reportero basado en fluorescencia para identificar los componentes celulares necesarios para la toxina de ricina una cadena (RTA) trata de levadura

Bacterial and plant A/B toxins exploit the natural trafficking pathways in eukaryotic cells to reach their intracellular target(s) in the cytosol and to ...

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Efecto de proteínas fluorescentes de fusión socios en ensayos de toxicidad de poliglutaminas de levadura

For the investigation of protein localization and trafficking using live cell imaging, researchers often rely on fusing their protein of interest to a ...