Name: isolation of human umbilical arterial smooth muscle cells (huasmc)
Uploaded: 2010-07-03T17:00:00
Description: Watch this Scientific Journal Video about Isolation of Human Umbilical Arterial Smooth Muscle Cells HUASMC at JoVE.com

<p class="jove_content"> Hospital Amato Lusitano, Castelo Branco Portugal. <br /> Hospital Sousa Martins, Guarda Portugal. </p>

<ol><li> Artérias são obtidos a partir de peças UC de 3-7 cm. </li><li> Geleia de Wharton que envolve as artérias foi cuidadosamente removido, cortando-a com uma tesoura. </li><li> Uma agulha sem corte-extremidade de um diâmetro externo de 0,6-1mm foi inserido cerca de 7 mm de uma extremidade das artérias dissecadas. </li><li> Para remover o sangue restante no interior dos vasos, as artérias são verticalmente realizada e perfundidos com, aproximadamente 20 a 40 mL, solução salina fisiológica (PSS), usando uma seringa de 20 ml conecta...

<ol>
	<li>Van Rijen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gap+junctions+in+human+umbilical+cord+endothelial+cells+contain+multiple+connexins.">Gap junctions in human umbilical cord endothelial cells contain multiple connexins.</a> <em style='font-style: italic'>The American journal of physiology</em>. <b>272</b> (1 PT 1), C117-C117 (1997).</li><li>Mart&#237;n de Llano, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Procedure+to+consistently+obtain+endothelial+and+smooth+muscle+cell+cultures+from+umbilical+cord+vessels.">Procedure to consistently obtain endothelial and smooth muscle cell cultures from umbilical cord vessels.</a> <em style='font-style: italic'>Translational Research</em>. <b>149</b> (1), 1-9 (2007).</li><li>Jaffe, E. A., Nachman, R. L., Becker, C. G., Minick, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culture+of+human+endothelial+cells+derived+from+umbilical+veins.+Identification+by+morphologic+and+immunologic+criteria.">Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria.</a> <em style='font-style: italic'>J Clin Invest</em>. <b>52</b> (11), 2745-2756 (1973).</li><li>Cairr&#227;o, E., Santos-Silva, A., Alvarez, E., Correia, I., Verde, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+culture+of+human+umbilical+artery+smooth+muscle+cells+expressing+functional+calcium+channels.">Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels.</a> <em style='font-style: italic'>In Vitro Cellular &#38; Developmental Biology-Animal</em>. <b>45</b> (3), 175-184 (2009).</li></ol>

<p class="jove_content"> O SMC pode ser usado como modelo para testes futuros com drogas constrictor, para isolar e caracterizar estruturalmente os canais de cálcio tipo L, para estudar os aspectos celulares e moleculares da função vascular e usá-los em engenharia de tecidos. SMCs e ECs são fundamentais para o desenvolvimento de novos biomateriais que podem ser usados ​​na produção de semi vasos sanguíneos sintético para ser usado em humanos. </p>

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>DMEM-F12</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>095K8311</td>
<td>&nbsp;</td>
<tr>
<td>Fetal bovine serum (FBS)</td>
<td>&nbsp;</td>
<td>Biochrom-AG</td>
<td>0117T</td>
<td>&nbsp;</td>
<tr>
<td>RPMI 1640</td>
<td>&nbsp;</td>
<td>Gibco</td>
<td>3103806</td>
<td>&nbsp;</td>
<tr>
<td>Collagen</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>067K2302</td>
<td>&nbsp;</td>
<tr>
<td>Antibiotic</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>10 &mu;L/ 1 mL of medium</td>
</tr>
<tr>
<td>Scissors</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Sterilized</td>
</tr>
<tr>
<td>Needle</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Diameter of 0.6-1mm</td>
</tr>
<tr>
<td>Syringe</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>10 - 20 mL</td>
</tr>
<tr>
<td>Physiological Saline Solution (PSS)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>20-40 mL per artery</td>
</tr>
<tr>
<td>Collagenase type I</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>047K1052</td>
<td>3 mL per artery</td>
</tr>
<tr>
<td>Hank's Buffered Salt Solution(HBSS)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Diluted collagenase </td>
</tr>
<tr>
<td>Phosphate buffered saline solution (PBS)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Warmed to 37&deg;C</td>
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isolation of human umbilical arterial smooth muscle cells (huasmc)

<p class="jove_content">O cordão umbilical humano (UC) é uma amostra biológica que pode ser facilmente obtida logo após o nascimento. Esta amostra biológica é, na maioria das vezes, descartada e sua coleção não implica qualquer risco acrescido para a saúde dos recém-nascidos ou a mãe s. Além disso há as questões éticas são levantadas. A UC é composta por uma veia e duas artérias do qual ambas as células endoteliais (CE)<sup&gt; 1</sup&gt; E células musculares lisas (PMEs)<sup&gt; 2</sup&gt;, Dois dos principais componentes celulares dos vasos sanguíneos, podem ser isoladas. Neste projeto o SMCs foram obtidos após o tratamento enzimático das artérias UC acordo com o procedimento experimental previamente descrito por Jaffe<em&gt; Et al</em<sup&gt; 3</sup&gt;. Após o isolamento de células que foram mantidos em t-flash com DMEM-F12 suplementado com 5% de soro fetal bovino e foram cultivadas por várias passagens. Células mantiveram sua morfologia e outras características fenotípicas nas gerações diferentes. O objetivo deste estudo foi isolar células musculares lisas, a fim de usá-los como modelos para ensaios futuros com drogas constrictor, isolar e caracterizar estruturalmente os canais de cálcio tipo L, para estudar os aspectos celulares e moleculares da função vascular<sup&gt; 4</sup&gt; E para usá-los em engenharia de tecidos.</p>

Watch this Scientific Journal Video about Isolation of Human Umbilical Arterial Smooth Muscle Cells HUASMC at JoVE.com

Isolation of Human Umbilical Arterial Smooth Muscle Cells HUASMC

<p class="jove_content">Os cordões umbilicais são usados ​​para isolar as células do músculo liso por caminhos diferentes. Neste trabalho utilizou-se o tratamento enzimático de células musculares lisas isoladas.</p>

Isolamento da Human Umbilical células musculares arteriais (HUASMC)

The human umbilical cord (UC) is a biological sample that can be easily obtained just after birth. This biological sample is, most of the time, discarded ...

Research

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Isolation of Human Umbilical Arterial Smooth Muscle Cells (HUASMC)

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