Name: isolation of primary patient-specific aortic smooth muscle cells and semiquantitative real-time contraction measurements in vitro
Uploaded: 2022-02-15T13:30:00
Description: Watch this Scientific Journal Video about Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro at JoVE.com

我们要感谢Tara van Merrienboer，Albert van Wijk，Jolanda van der Velden，Jan D. Blankensteijn，Lan Tran，Peter L. Hordijk，PAREL-AAA团队以及阿姆斯特丹UMC，Zaans Medisch Centrum和Dijklander医院的所有血管外科医生为这项研究提供了材料和支持。

注意：在阿姆斯特丹大学医学中心，阿姆斯特丹VU大学医学中心，Zaans Medisch Centrum，Zaandam和荷兰霍恩的Dijklander医院的开放动脉瘤修复期间获得了主动脉瘤活检。控制主动脉组织是从附着在肾动脉上的主动脉碎片获得的，用于肾移植。仅纳入18岁以上的患者，所有患者都表示知情同意参加该研究。所有材料均按照赫尔辛基WMA宣言的规定和VU医疗中心医学伦理委员会的机构指南收集。所有实验和实验?...

<ol>
	<li>Milewicz, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+basis+of+thoracic+aortic+aneurysms+and+dissections:+focus+on+smooth+muscle+cell+contractile+dysfunction.">Genetic basis of thoracic aortic aneurysms and dissections: focus on smooth muscle cell contractile dysfunction.</a> Annual Review of Genomics and Human Genetics. 9, 283-302 (2008).</li><li>Milewicz, D. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+smooth+muscle+cell+force+generation+as+a+driver+of+thoracic+aortic+aneurysms+and+dissections.">Altered smooth muscle cell force generation as a driver of thoracic aortic aneurysms and dissections.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 37 (1), 26-34 (2017).</li><li>Groeneveld, M. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Betaglycan+(TGFBR3)+up-regulation+correlates+with+increased+TGF-&#946;+signaling+in+Marfan+patient+fibroblasts+in+vitro.">Betaglycan (TGFBR3) up-regulation correlates with increased TGF-&#946; signaling in Marfan patient fibroblasts in vitro.</a> Cardiovascular Pathology. 32, 44-49 (2018).</li><li>Chen, J., Li, H., SundarRaj, N., Wang, J. H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha-smooth+muscle+actin+expression+enhances+cell+traction+force.">Alpha-smooth muscle actin expression enhances cell traction force.</a> Cell Motility and the Cytoskeleton. 64 (4), 248-257 (2007).</li><li>Peyton, S. R., Putnam, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+matrix+rigidity+governs+smooth+muscle+cell+motility+in+a+biphasic+fashion.">Extracellular matrix rigidity governs smooth muscle cell motility in a biphasic fashion.</a> Journal of Cellular Physiology. 204 (1), 198-209 (2005).</li><li>Williams, D. A., Fogarty, K. E., Tsien, R. Y., Fay, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+gradients+in+single+smooth+muscle+cells+revealed+by+the+digital+imaging+microscope+using+Fura-2.">Calcium gradients in single smooth muscle cells revealed by the digital imaging microscope using Fura-2.</a> Nature. 318 (6046), 558-561 (1985).</li><li>Wu, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NLRP3+(nucleotide+oligomerization+domain-like+receptor+family,+pyrin+domain+containing+3)-caspase-1+inflammasome+degrades+contractile+proteins:+implications+for+aortic+biomechanical+dysfunction+and+aneurysm+and+dissection+formation.">NLRP3 (nucleotide oligomerization domain-like receptor family, pyrin domain containing 3)-caspase-1 inflammasome degrades contractile proteins: implications for aortic biomechanical dysfunction and aneurysm and dissection formation.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 37 (4), 694-706 (2017).</li><li>Bogunovic, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+smooth+muscle+cell+contractility+as+a+novel+concept+of+abdominal+aortic+aneurysm+pathophysiology.">Impaired smooth muscle cell contractility as a novel concept of abdominal aortic aneurysm pathophysiology.</a> Scientific Reports. 9 (1), 1-14 (2019).</li><li>Hurst, V., Goldberg, P. L., Minnear, F. L., Heimark, R. L., Vincent, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rearrangement+of+adherens+junctions+by+transforming+growth+factor-&#946;1:+role+of+contraction.">Rearrangement of adherens junctions by transforming growth factor-&#946;1: role of contraction.</a> American Journal of Physiology-Lung Cellular and Molecular Physiology. 276 (4), 582-595 (1999).</li><li>Hu, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+between+ECIS+and+LAPS+for+establishing+a+cardiomyocyte-based+biosensor.">Comparison between ECIS and LAPS for establishing a cardiomyocyte-based biosensor.</a> Sensors and Actuators B: Chemical. 185, 238-244 (2013).</li><li>Peters, M. F., Lamore, S. D., Guo, L., Scott, C. W., Kolaja, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+stem+cell-derived+cardiomyocytes+in+cellular+impedance+assays:+bringing+cardiotoxicity+screening+to+the+front+line.">Human stem cell-derived cardiomyocytes in cellular impedance assays: bringing cardiotoxicity screening to the front line.</a> Cardiovascular Toxicology. 15 (2), 127-139 (2015).</li><li>Zhang, S., Yang, Y., Kone, B. C., Allen, J. C., Kahn, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin-stimulated+cyclic+guanosine+monophosphate+inhibits+vascular+smooth+muscle+cell+migration+by+inhibiting+Ca/calmodulin-dependent+protein+kinase+II.">Insulin-stimulated cyclic guanosine monophosphate inhibits vascular smooth muscle cell migration by inhibiting Ca/calmodulin-dependent protein kinase II.</a> Circulation. 107 (11), 1539-1544 (2003).</li><li>Halterman, J. A., Kwon, H. M., Zargham, R., Bortz, P. D. S., Wamhoff, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+factor+of+activated+T+cells+5+regulates+vascular+smooth+muscle+cell+phenotypic+modulation.">Nuclear factor of activated T cells 5 regulates vascular smooth muscle cell phenotypic modulation.</a> Arteriosclerosis, Thrombosis, and Vascular Biology. 31 (10), 2287-2296 (2011).</li><li>Bass, H. M., Beard, R. S., Cha, B. J., Yuan, S. Y., Nelson, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thrombomodulin+induces+a+quiescent+phenotype+and+inhibits+migration+in+vascular+smooth+muscle+cells+in+vitro.">Thrombomodulin induces a quiescent phenotype and inhibits migration in vascular smooth muscle cells in vitro.</a> Annals of Vascular Surgery. 30, 149-156 (2016).</li><li>Burger, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+phenotyping+and+functional+assessment+of+smooth+muscle+like-cells+with+pathogenic+variants+in+aneurysm+genes+ACTA2,+MYH11,+SMAD3+and+FBN1.">Molecular phenotyping and functional assessment of smooth muscle like-cells with pathogenic variants in aneurysm genes ACTA2, MYH11, SMAD3 and FBN1.</a> Human Molecular Genetics. , (2021).</li><li>Yeung, K. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transdifferentiation+of+human+dermal+fibroblasts+to+smooth+muscle-like+cells+to+study+the+effect+of+MYH11+and+ACTA2+mutations+in+aortic+aneurysms.">Transdifferentiation of human dermal fibroblasts to smooth muscle-like cells to study the effect of MYH11 and ACTA2 mutations in aortic aneurysms.</a> Human Mutation. 38 (4), 439-450 (2017).</li></ol>

本文提出了一种基于阻抗和表面占用变化 的体外测量SMC收缩的方法。首先，描述了患者特异性原代人类SMC和皮肤成纤维细胞的分离，培养和扩增，然后描述了如何使用它们进行收缩测量。 
该研究的局限性与通过外植体方案获得细胞有关。从活检中增殖的细胞可能具有与 体内原始组织不同的性质。此外，这里介绍的外植体方案并不具有创新性，但包括用于提供?...

平滑肌细胞（SMC）是主动脉内侧层中的主要细胞类型，是主动脉最厚的一层。在壁内，它们是径向的，除其他功能外，还参与血管收缩和血管舒张1。SMC收缩机制通过与细胞外基质2的功能连接参与主动脉中力的传递。编码SMC收缩器官蛋白质的基因突变，如平滑肌肌球蛋白重链（MYH11）和平滑肌肌动蛋白（ACTA2），与家族性胸主动脉瘤病例有关，强调了SMC收缩在维持主动脉结构和功能完整性方面的相关性1，2.此外，TGFβ信号通路中的突变也与主动脉瘤有关，它们在主动脉瘤病理生理学中的作用也可以在皮肤成纤维细胞中研究3。
体外SMC收缩的高通量测量具有挑战性。由于SMC收缩力不能在人体体内测量，因此在人体细胞上进行体外测定是一种可行的替代方案。此外，动物模型中的腹主动脉瘤（AAA）发展要么是由弹性蛋白酶灌注等化学诱导的，要么是由特定突变引起的。因此，动物数据不能与人类的AAA发展相提并论，后者大多具有多因素原因，例如吸烟，年龄和/或动脉粥样硬化。体外迄今为止，SMC收缩力主要通过牵引力显微镜4，5，荧光细胞内钙通量6的定量和胶原皱纹测定7来测定。虽然牵引力显微镜提供了对单个细胞产生的力的宝贵数字洞察力，但由于复杂的数学数据处理和一次分析一个细胞，它不适合高通量筛选，这意味着测量每个供体的代表性细胞数量非常耗时。Fura-2染料和胶原起皱测定允许表面上确定收缩，并且不能给出精确的数值输出，使它们不太适合区分患者特异性差异。通过优化一种测量体外SMC收缩的新方法，首次证明了腹主动脉瘤患者主动脉瘤患者主动脉瘤细胞中SMC收缩受损8。这是通过重新利用电电池基板阻抗传感（ECIS）方法完成的。ECIS是一种实时的中等通量测定法，用于定量贴壁细胞行为和收缩9，10，11，例如SMC生长和伤口愈合和迁移测定中的行为12，13，14。确切的方法在协议部分进行了描述。以这种优化的方式，ECIS也可用于研究成纤维细胞收缩，因为它们的大小和形态相似。
本文的目的是逐步描述使用ECIS8测量体外SMC收缩的方法，并比较对照组和患者SMC之间的收缩。首先，解释了原代SMC从对照和患者主动脉活检中的分离和培养，可用于收缩测量。其次，描述了收缩测量和分析，以及SMC标志物表达的验证。此外，本文描述了分离患者特异性真皮成纤维细胞的方法，其收缩可以使用相同的方法测量。这些细胞可用于专注于主动脉瘤或其他心血管病理学的患者特异性研究15，或使用转分化方案的预后研究，其允许在动脉瘤手术前进行收缩测量16。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>96-well Array</td>
			<td>Applied Biophysics</td>
			<td>96W10idf PET</td>
			<td>Array used to measure contraction in the ECIS setup</td>
		</tr>
		<tr>
			<td>Custodiol</td>
			<td>Dr. Franz H&#246;hler Chemie GmbH</td>
			<td>RVG 12801</td>
			<td>Solution used to transfer tissue in from surgery room to laboratorium</td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide</td>
			<td>Sigma-Aldrich</td>
			<td>472301</td>
			<td>Solution used to dilute ionomycin</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Gibco</td>
			<td>26140079</td>
			<td>Addition to cell culture medium</td>
		</tr>
		<tr>
			<td>Ham&#39;s F-10 Nutrient Mix</td>
			<td>Gibco</td>
			<td>11550043</td>
			<td>Medium used to culture skin fibroblasts</td>
		</tr>
		<tr>
			<td>Human Vascular Smooth Muscle Cell Basal Medium (formerly &#39;&#39;Medium 231&#39;&#39;)</td>
			<td>Gibco</td>
			<td>M231500</td>
			<td>Medium used to culture smooth muscle cells</td>
		</tr>
		<tr>
			<td>Invitrogen countess II</td>
			<td>Thermo Fisher Scientific</td>
			<td>AMQAX1000</td>
			<td>Automated cell counter</td>
		</tr>
		<tr>
			<td>Ionomycin calcium salt from Streptomyces conglobatus</td>
			<td>Sigma-Aldrich</td>
			<td>I0634-1MG</td>
			<td>Compound used for contraction stimulation</td>
		</tr>
		<tr>
			<td>NaCl 0.9%</td>
			<td>Fresenius Kabi</td>
			<td>B230561</td>
			<td>Solution used to transfer tissue in from surgery room to laboratorium</td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin</td>
			<td>Gibco</td>
			<td>15140122</td>
			<td>Antibiotics used for cell culture medium</td>
		</tr>
		<tr>
			<td>Phospathe buffered saline</td>
			<td>Gibco</td>
			<td>10010023</td>
			<td>Used to wash cells</td>
		</tr>
		<tr>
			<td>Quick-RNA Miniprep Kit</td>
			<td>Zymo Research</td>
			<td>R1055</td>
			<td>Kit used for RNA isolation</td>
		</tr>
		<tr>
			<td>Smooth Muscle Growth Supplement (SMGS)</td>
			<td>Gibco</td>
			<td>S00725</td>
			<td>Supplement which is added to smooth muscle cell culture medium</td>
		</tr>
		<tr>
			<td>SuperScript VILO cDNA Synthesis Kit</td>
			<td>Thermo Fisher Scientific</td>
			<td>11754250</td>
			<td>Kit used for cDNA synthesis</td>
		</tr>
		<tr>
			<td>SYBR Green PCR Master Mix</td>
			<td>Thermo Fisher Scientific</td>
			<td>4309155</td>
			<td>Reagent for qPCR</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA</td>
			<td>Gibco</td>
			<td>15400-054</td>
			<td>Used to trypsinize cells</td>
		</tr>
		<tr>
			<td>ZTheta</td>
			<td>Applied Biophysics</td>
			<td>ZTheta</td>
			<td>ECIS instrument used for contraction measurements</td>
		</tr>
	</tbody>
</table>

isolation of primary patient-specific aortic smooth muscle cells and semiquantitative real-time contraction measurements in vitro

平滑肌细胞（SMC）是主动脉介质中的主要细胞类型。它们的收缩机制对于主动脉中力的传递很重要，并调节血管收缩和血管舒张。编码SMC收缩器官蛋白的基因突变与主动脉疾病有关，例如胸主动脉瘤。 在体外 测量SMC收缩具有挑战性，特别是以高通量方式，这对于筛查患者材料至关重要。当前可用的方法不适合此目的。本文提出了一种基于电电池基板阻抗传感（ECIS）的新方法。首先，描述了一种外植体方案，用于从主动脉活检和患者特异性人原发性真皮成纤维细胞中分离出患者特异性人原发性SMC，以研究主动脉瘤。接下来，详细描述了一种新的收缩方法来测量这些细胞的收缩反应，包括随后的分析和比较不同组的建议。该方法可用于在转化（心血管）研究以及患者和药物筛选研究的背景下研究贴壁细胞的收缩。

为了测试该方法的可重复性，该方法首先仅使用对照SMC进行验证。为了确定实验间测量的再现性，将所有纳入的对照和患者细胞系的两个独立测量绘制为Bland-Altman图（图3B）。该图表明，除了一个异常值细胞系外，该方法在置信区间之外没有显示出变异性。此外，这些结果表明，在同一实验中播种并同时刺激的两个孔显示出几乎相同的收缩反应曲线（图4C</stro...

Watch this Scientific Journal Video about Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro at JoVE.com

Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro

本文介绍了一种基于外植体培养的方法，用于分离和培养原代患者特异性人主动脉平滑肌细胞和真皮成纤维细胞。此外，还提出了一种用于测量细胞收缩和后续分析的新方法，可用于研究这些细胞中患者特异性差异。

原发性患者特异性主动脉平滑肌细胞的分离和体外半定量实时收缩测量

Smooth muscle cells (SMCs) are the predominant cell type in the aortic media. Their contractile machinery is important for the transmission of force in ...

这种方法允许研究人员同时研究多个患者细胞系的细胞收缩。它使我们能够更快，更有效地研究疾病。这种技术使我们能够实时量化细胞收缩。在一小时内，我们可以获得数十个单元格的收缩值。我们首次证明，在一组主动脉瘤患者中，肌肉细胞收缩减少。这可能是一种新的生物标志物或药物治疗的靶点。首先对两对手术镊子和手术刀进行消毒，将它们浸入70%乙醇中，然后将其擦干。然后在培养皿中移液两毫升SMC培养基，其中将进行组织解剖。接下来将2.5毫升SMC培养基移液到两个T-25烧瓶中。旋转烧瓶，使少量的培养基覆盖整个表面。目视检查活检，通过内膜侧存在动脉粥样硬化斑块和外侧存在粘稠结缔组织来识别三个主动脉层，以区分这些层。为了从培养基中分离SMC，首先将具有内膜斑块的组织侧朝上放置，然后通过用镊子将其从组织中拉出来除去固体斑块，同时用第二对镊子向下推组织，直到粉红色均匀的内侧层可见。现在翻转组织并通过拉下外膜层重复相同的过程，确保根据需要多次尝试移除所有可见部分，因为该层不会轻易从介质中分离。一旦介质层被分离，通过用镊子将培养基压下，然后将组织切成立方体，并使用手术刀在一个方向上切割组织，使清洁的单向切割以尽量减少损伤。使用镊子将10至20个这些组织碎片放在T-25弹片的上四分之一处。如前所述，在对两对手术镊子和手术刀进行灭菌后，在培养皿中移液两毫升成纤维细胞培养基，其中将进行组织解剖。然后将2.5毫升成纤维细胞培养基移液到两个T-25烧瓶中，并旋转烧瓶，使少量培养基覆盖整个表面。目视检查活检以识别具有可识别皮肤表面的表皮，有时具有可见的头发，然后在另一侧寻找黄色和粘稠的皮下脂肪。表皮和皮下脂肪之间的层是真皮，是活成纤维细胞的来源。为了从真皮层中分离成纤维细胞，通过将组织放在真皮层一侧来去除其他两层，以使所有三层都可见。然后用镊子按住组织，并在一条干净的线上切除整个表皮，而不会损坏组织。现在翻转组织并重复相同的过程，方法是在与皮下脂肪平行的真皮内切割。一旦真皮被分离，将组织切成立方体，用镊子将组织压下，然后使用手术刀将组织切成一个方向。然后使用镊子将10至20个这些组织片放入T-25烧瓶的上四分之一。使用自动细胞计数器计数细胞，并在明胶包被的ECIS板中以每孔30， 000个细胞的接种密度在200微升SMC培养基中一式三份接种SMC。将板与SMC一起放入细胞培养箱中的ECIS的96孔支架中。双击 ECIS 应用生物物理学软件以打开程序，然后按“设置”按钮。检查所有电极是否都与左侧下面板中标有“孔配置”的支架接触。如果电极连接不正确，请在开始测量之前调整支架中的板。现在，通过单击阵列类型在同一面板中选择板类型。接下来，准备两个液管，设置为两微升和150微升体积。在开始刺激之前，按软件中的标记并发表评论。为了刺激细胞，取下盖子并将其放在培养箱内的无菌表面上。然后通过将两微升的离子霉素工作溶液移液到每个孔中来诱导SMC收缩。一旦所有细胞都受到刺激，使用第二个移液管在孔中混合培养基。为了确定实验间测量的再现性，将所有纳入的对照和患者细胞系的两个独立测量绘制为Bland-Altman图，证明该方法除了一个异常值细胞系外，没有显示出置信区间外的变异性。用同一实验播种并同时刺激的两个孔显示出几乎相同的契约反应曲线。为了验证所显示的反应是否由于离子霉素刺激而收缩而不是渗透应激，在刺激后更换培养基并记录细胞的行为，这表明受刺激的细胞再次开始在电极上扩散。来自健康、非扩张主动脉的SMC的收缩反应显示中位反应为61%，而腹主动脉瘤患者的中位反应为52%。对照组的平均收缩反应用于识别4名收缩值低于正常值的患者。蛋白质印迹分析和随后的定量证实细胞表达SMC标志物。为了确定SMC的增殖能力，对Ki67进行了qPCR。Ki67表达在所有细胞中均可检测到，但与收缩输出不相关。当刺激细胞收缩时，请记住不要同时移液过多的孔，并尽量尽快移液。此过程可能需要一些练习。使用这种方法，我们根据患者的收缩和降低正常收缩来划分患者，这使我们能够研究这些患者及其临床特征（如吸烟）的联系。

Research

JoVE Journal

Biology

Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro

Isolation of Human Umbilical Arterial Smooth Muscle Cells (HUASMC)

Isolation of Human Umbilical Arterial Smooth Muscle Cells HUASMC

人脐动脉平滑肌细胞的分离（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 ...

科研

生物学

Isolation and Primary Culture of Rat Hepatic Cells

大鼠肝细胞的分离和小学文化

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

Isolation of Pulmonary Artery Smooth Muscle Cells from Neonatal Mice

肺动脉平滑肌细胞从新生小鼠隔离

Pulmonary hypertension is a significant cause of morbidity and mortality in infants. Historically, there has been significant study of the signaling ...

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance (SPR)

Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR

膜蛋白的实时测量：受体相互作用使用表面等离子体共振（SPR）

Protein-protein interactions are pivotal to most, if not all, physiological processes, and  understanding the nature of such interactions is a central ...

Isolation and Primary Culture of Mouse Aortic Endothelial Cells

隔离和鼠标主动脉内皮细胞原代培养

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an ...

Isolation of Murine Coronary Vascular Smooth Muscle Cells

鼠冠状动脉血管平滑肌细胞的分离

While the isolation and culture of vascular smooth muscle cells (VSMCs) from large vessels is well established, we sought to isolate and culture VSMCs ...

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

小鼠主动脉夹层损伤：高效<em&gt;体内</em&gt;平滑肌细胞增殖和内皮功能模型

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell ...

Isolation and Culture of Primary Aortic Endothelial Cells from Miniature Pigs

微型猪原主主动脉内皮细胞的隔离与培养

Xenotransplantation is a promising way to resolve the shortage of human organs for patients with end-stage organ failure, and the pig is considered as a ...

Measuring Proliferation of Vascular Smooth Muscle Cells Using Click Chemistry

使用点击化学测量血管平滑肌细胞增殖

The ability of a cell to proliferate is integral to the normal function of most cells, and dysregulation of proliferation is at the heart of many disease ...

Isolation of Mouse Interstitial Valve Cells to Study the Calcification of the Aortic Valve In Vitro

Isolation of Mouse Interstitial Valve Cells to Study the Calcification of the Aortic Valve In Vitro

小鼠间质瓣细胞分离研究主动脉瓣体外钙化

The calcification of aortic valve cells is the hallmark of aortic stenosis and is associated with valve cusp fibrosis. Valve interstitial cells (VICs) ...