Name: primary culture of adult rat heart myocytes
Uploaded: 2009-06-16T00:00:00
Description: Watch this Scientific Journal Video about Primary Culture of Adult Rat Heart Myocytes at JoVE.com

<p class="jove_title">部分一，准备进行手术</p><ol><li>大鼠手术的前一天，确信所有的解决方案，但不添加任何酶。缓冲器A，B和洗涤缓冲液都应该被过滤为不育之前存储。缓冲区都应该保存在4 ° C </li><li> 10-20微克/毫升层粘连蛋白在2 mL 1XSFM以及前一天必须镀有6孔组织培养板，并应存放在37 ° C中的CO <sub>2</sub>培养箱。 </li><li>在手术当天，准备用70％乙醇消毒钳，剪刀，镊子，手术器械，并让他们干纸巾。 </li><li>充满0.5 ...

<ol>
	<li>Miriyala, J., Nguyen, T., Yue, D. T., Colecraft, H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+CaVbeta+subunits,+and+lack+of+functional+reserve,+in+protein+kinase+A+modulation+of+cardiac+CaV1.2+channels.">Role of CaVbeta subunits, and lack of functional reserve, in protein kinase A modulation of cardiac CaV1.2 channels.</a> <em style='font-style: italic'>Circ Res</em>. <b>102</b> (7), e54-e64 (2008).</li><li>SX, T. a. k. a. h. a. s. h. i., Mittman, S., Colecraft, H. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinctive+modulatory+effects+of+five+human+auxiliary+beta2+subunit+splice+variants+on+L-type+calcium+channel+gating.">Distinctive modulatory effects of five human auxiliary beta2 subunit splice variants on L-type calcium channel gating.</a> <em style='font-style: italic'>Biophys</em>. , 84-845 (2003).</li></ol>

<p class="jove_content">关键的一步，这是挂在离体心脏灌注系统的速度。酶消化期的长短可能会有点不同的各组大鼠。调整取决于软心定期的消化期后成为。 “缓慢复苏的Ca <sup>2 +</sup>内切酶消化后获得的Ca <sup>2 +</sup>宽容健康的细胞至关重要。 </p><p class="jove_content">豚鼠，该协议是相同的，除了透明质酸是第十四蛋白酶，而不是使用。通常情况下，我们发现，最初的活细胞百分比是豚鼠至大鼠相比，虽然他们生存，仅...

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		<th>Type</th>
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		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>NaCl</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>S6191</td>
<td>&nbsp;</td>
<tr>
<td>KCl</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P9541</td>
<td>&nbsp;</td>
<tr>
<td>HEPES</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>H4034</td>
<td>&nbsp;</td>
<tr>
<td>K<sub>2</sub>HPO<sub>4</sub></td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P9666</td>
<td>&nbsp;</td>
<tr>
<td>MgSO<sub>4</sub></td>
<td>&nbsp;</td>
<td>Fluka</td>
<td>63068</td>
<td>&nbsp;</td>
<tr>
<td>Glucose</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>G7021</td>
<td>&nbsp;</td>
<tr>
<td>CaCl<sub>2</sub></td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>C7902</td>
<td>&nbsp;</td>
<tr>
<td>Heparin Sodium Salt</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>H4784</td>
<td>&nbsp;</td>
<tr>
<td>Collagenase Type II</td>
<td>&nbsp;</td>
<td>Worthington Biochemical</td>
<td>LS004176</td>
<td>&nbsp;</td>
<tr>
<td>Protease XIV</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P5147</td>
<td>&nbsp;</td>
<tr>
<td>2,3-Butanedione Monoxime (BDM)</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>B0753</td>
<td>&nbsp;</td>
<tr>
<td>Carnitine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>C9500</td>
<td>&nbsp;</td>
<tr>
<td>Taurine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>T0625</td>
<td>&nbsp;</td>
<tr>
<td>Glutamic Acid</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>G1501</td>
<td>&nbsp;</td>
<tr>
<td>BSA</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>A3294</td>
<td>&nbsp;</td>
<tr>
<td>Creatine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>C3630</td>
<td>&nbsp;</td>
<tr>
<td>Antibiotic</td>
<td>&nbsp;</td>
<td>Cellgro</td>
<td>30-009-CI</td>
<td>&nbsp;</td>
<tr>
<td>Media 199</td>
<td>&nbsp;</td>
<td>GIBCO</td>
<td>11150</td>
<td>&nbsp;</td>
<tr>
<td>FBS</td>
<td>&nbsp;</td>
<td>Hyclone</td>
<td>SH30071.03</td>
<td>&nbsp;</td>
<tr>
<td>Laminin</td>
<td>&nbsp;</td>
<td>BD Bioscience</td>
<td>354232</td>
<td>&nbsp;</td>
<tr>
<td>water bath</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>15-462-5</td>
<td>&nbsp;</td>
<tr>
<td>variable flow chemical pump</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>15-077-67</td>
<td>&nbsp;</td>
<tr>
<td>6-well culture plate</td>
<td>&nbsp;</td>
<td>Corning</td>
<td>3516</td>
<td>&nbsp;</td>		</tbody>
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primary culture of adult rat heart myocytes

<p class="jove_content">原代培养成年啮齿类动物的心脏细胞，心血管研究的一个重要的模型系统。不过，建立强大的，可行的的培养成人心肌细胞可以在技术上具有挑战性的，速率限制，许多研究者的一步。在这里，我们描述了一个协议，以获得一个可行的几天停留在文化的成年大鼠心脏心肌细胞的高产。心是孤立的，与胶原酶和蛋白酶灌注下低钙<sup&gt; 2 +</sup&gt;恢复单个心肌细胞的条件。钙<sup&gt; 2 +</sup&gt;容错细胞得到逐步增加细胞外Ca<sup&gt; 2 +</sup&gt;集中在三个随后的清洗步骤。被过滤，在培养液中悬浮细胞，并镀金层粘连蛋白涂单。使用此协议培养的心肌细胞获得长达4天的可行和适合最，包括电生理学，生物化学，影像和分子生物学的实验。</p>

Watch this Scientific Journal Video about Primary Culture of Adult Rat Heart Myocytes at JoVE.com

Primary Culture of Adult Rat Heart Myocytes

<p class="jove_content">在本文中，我们描述了一个典型的成年大鼠心脏心肌细胞分离和培养。胶原酶和蛋白酶消化，分离单个心肌细胞。心肌细胞培养遵守此协议满足大多数实验要求。</p>

成年大鼠心脏心肌细胞的原代培养

Cultured primary adult rodent heart cells are an important model system for cardiovascular research.  Nevertheless, establishment of robust, viable ...

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The function of mitochondria in generation of cellular ATP in the process of oxidative phosphorylation is widely recognised. During the past decades there ...

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Primary hepatocyte culture is a valuable tool that has been extensively used in basic research of liver function, disease, pathophysiology, pharmacology ...

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We identified a novel population of melanocyte-like cells (also known as cardiac melanocytes) in the hearts of mice and humans that contribute to atrial ...

Methods for the Isolation, Culture, and Functional Characterization of Sinoatrial Node Myocytes from Adult Mice

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Sinoatrial node myocytes (SAMs) act as the natural pacemakers of the heart, initiating each heart beat by generating spontaneous action potentials (APs). ...

Simultaneous Isolation of High Quality Cardiomyocytes, Endothelial Cells, and Fibroblasts from an Adult Rat Heart

从成年大鼠心脏同时分离高质量心肌细胞，内皮细胞和成纤维细胞

The rat is an important animal model used in cardiovascular research, and rat cardiac cells are used routinely for in vitro analysis of the molecular ...

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The keratinocyte (KC) is the predominant cell type in the epidermis, the outermost layer of the skin. Epidermal KCs play a critical role in providing skin ...