Name: the inverted heart model for interstitial transudate collection from the isolated rat heart
Uploaded: 2017-06-20T15:00:00.000+00:00
Duration: 7 min 59 s
Description: Watch this Scientific Journal Video about The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart at JoVE.com

本研究由NSFC 81570244，FoKo 23/2013和SFB 1116 / B01以及杜塞尔多夫心血管研究所（CARID）资助。

所有实验均经当地监管机构（德国Nordrhein-Westfalen的LANUV）批准，并按照动物使用指南进行。动物喂食标准食物， 随意自来水。实验每个步骤所需的所有设备和化学品均可在材料表中找到 。 1.准备乳胶帽和室内气球<ol><li>使用与大鼠心脏平均体重（体重300-350克）匹配的铣床制作铝模具。用超细（10/0）砂纸抛光模具。 注意:模具的详细指标如图1A所示。 </li><li>将铝模具的颈部垂直固定到铣床上以制备胶乳盖。 注意:铣床使模具缓慢旋转。或者，可以使用电动机。 </<html> LI&gt; <li>将20mL液体胶乳（商业购买，参见材料表 ）倒入50mL玻璃烧杯中。 </li><li>降低模具直到模具的整个身体浸入胶乳溶液中。 </li><li>在旋转的同时慢慢抬起模具（5厘米/分钟）。 </li><li>继续旋转模具15分钟，直到模具表面的胶乳固化。 </li><li>在模具的表面（已经被薄的胶乳膜覆盖）上加入约1g滑石粉，以防止在分离时损坏。 </li><li>用手指从模具表面轻轻取下已经干燥的乳胶帽;胶乳帽现在可以使用（ 图1B ）。 </li><li>将胶乳帽的出口连接到15厘米硅胶管（ID = 0.2毫米），以后用于收集CT。 </li><li>用水填充心室乳胶球囊并将其牢固地固定在连接到1毫升充水注射器的L形金属插管（&gt;图1C）。 注意:这将用于确保心脏的直立定位（见下文）。 </li><li>使用附带的1 mL注射器进行几次放气/充气试验，确保气囊密封。 </li><li>将插管通过三通止动器连接到压力传感器，以便将来测量心室内发育压力（ 图1C ）。 </li></ol> 2.制备Krebs-Henseleit缓冲液（KHB）和Langendorff灌注系统<ol><li>通过使用恒流（由辊泵驱动）或恒压（由玻璃柱中的静压产生）模式来设置Langendorff灌注系统。 注意:Langendorff心脏准备的细节已经在之前描述过12 。 </li><li> 制备2L改性KHB（mM:116.02NaCl，4.63KCl，1.10 MgSO 4 ·7H 2 O，1.21K 2 HPO 4 2 ·2H 2 O，24.88 NaHCO 3，8.30 D-葡萄糖和2.0丙酮酸钠）。 <ol><li>将所有化学品称重，然后称取CaCl 2，并将其溶解于2升烧瓶中1.8升双蒸水中。 </li><li>在磁力搅拌下，将含有碳原子（95％O 2 /5％CO 2 ）的培养基鼓泡至少5分钟以平衡（pH:7.4）。 </li><li>加入0.74g CaCl 2· 2H 2 O，并用双蒸水将总体积达到2L。 </li><li>继续搅拌并用碳源鼓泡培养基5分钟。 </li><li>通过0.2μm过滤器过滤KHB，以消除可能阻碍心脏微循环的小颗粒。 </li></ol></li><li> Langendorff灌注系统的准备。 <ol><li>将过滤的KHB置于预热水浴（38℃）中;保持鼓泡与碳源产生100 cmH 2 O的内部压力KHB水库。 </li><li>将储存器连接到玻璃柱，以建立用于KHB的Langendorff灌注的100cmH 2 O静水压力;继续用碳源鼓泡柱内的KHB。 </li><li>调整加温系统的温度，使主动脉插管出口处的温度为37°C。 </li><li>确保油管系统无气泡。 </li><li>将KHB与碳源再氧化5分钟，直到KHB中的PO 2达到500-600mmHg（通过血气分析仪测量）。 </li></ol></li><li>将灌注系统设置为在100 cmH 2 O的恒定压力下运行，或者使用手动切换以约10-20 mL / min的恒定流量运行。或者，使用可互换的STH泵控制器立即切换到灌注模式。 </li></ol>心脏的隔离和插管注意:体重为30的雄性Wistar大鼠使用0-350g，使得心脏的尺寸与预制乳胶帽匹配。大鼠进行左侧动脉下垂（LAD）的连接50分钟，然后再灌注或假手术。报告了心肌梗死（MI）诱导方法的细节。手术后5天对梗死动物进行逆心实验。 <ol><li>使用连接到动物饲养室（20L）的异氟烷蒸发器（2％V / V）麻醉大鼠。 </li><li>深度麻醉后，将大鼠转移到手术台（不受温度控制）。 </li><li>使用镊子将肌肉和肌肉提起到胸骨正下方，用重型剪刀将肋骨的下边缘切开。 </li><li>使用精细的剪刀，在肋边缘做一个小切口的隔膜。尾部切割，使整个腹侧胸壁的瓣。 </li><li>用拇指轻轻抓住心脏索引和中指，并缓慢向上提升，使心脏容器变得轻微拉伸。 </li><li>消除心脏直到主动脉完全暴露。 </li><li>将心脏置于含有50mL冰冷KHB（4℃）的100mL烧杯中，并将其移至灌注装置。 </li><li>立即将心脏通过主动脉安装到滴水插管上，并用缝合线（4-0）牢固地拧紧。避免气泡进入心脏。 </li><li>施加恒定灌注压力（100 cmH 2 O）。或者，可以应用完全流速（以20mL / min开始）。 注意:从胸部开放到心脏附着到灌注套管的时间应该在有经验的操作者手中约3分钟。 </li></ol>反转心脏模型<ol><li>轻轻旋转主动脉插管，直到心脏后壁处于正面 。 </li><li>用剪刀取出结缔组织暴露左心房的开口，使其准备好用于心室内插管。 </li><li>将通过左心房连接到刚性导管的放气乳胶球囊插入左心室。 </li><li>充气气球，直到充满整个心室（充气体积被预先标记在注射器上）。 </li><li>反转心脏直到它倒置，由心室内气囊导管支持它。 </li><li> 如图1C所示 ，使用具有刚性金属导管的心室内球囊将反转心脏机械稳定在直立位置。 </li><li>调整心脏的位置以避免主动脉根部的过度扭曲。 </li><li>将舒张压调整至3-5 mmHg（由室内球囊测量; 见图1C ）。 </li><li>观察心脏的心外膜表面，确保形成小液滴。 </li><li>放乳胶帽o通过轻轻推动心脏的表面，用手指覆盖整个心脏。 </li><li>确保乳胶帽覆盖大部分心室表面。 </li><li>通过用1-mL注射器轻轻吸取，去除盖子和管道内的气泡（如果有的话）。 </li><li>将CT放气管的远端开口调整到心脏水平面以下10厘米处。 注意:此步骤确保通过负静压力轻微吸吮。 </li><li>在1.5mL收集管中收集CT滴，放入冰中，与NaCl混合1:1。收集约0.15-1.5 mL的CT。 注意:冰/ NaCl混合物将采集管中的温度稳定在零下（约-4°C）。 注意:取样时间取决于实验目的。假手术动物（n = 3），冠状动脉结扎动物（n = 11）的CT流量约为27±20μL/ min。 </li><li>称重和快速冻结CT样品在液氮中并将其储存在-80°C进行后续测量。 </li></ol> 5. CT分析<ol><li>根据科学问题，使用CT流体分析代谢物。 注意:从恒压灌注（100cmH 2 O）收集图2和图3所示的数据，并在10分钟内收集约0.15-1.5mL CT流体。这个时间和体积对于各种嘌呤的蛋白质组学（最小值:50μL; 图2 ） 14和HPLC（最小值:20μL; 图3 ） 15是足够的。 </li></ol>

<ol>
	<li>Henkel, D. M., Redfield, M. M., Weston, S. A., Gerber, Y., Roger, V. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Death+in+heart+failure:+a+community+perspective.">Death in heart failure: a community perspective.</a> Circ Heart Fail. 1 (2), 91-97 (2008).</li><li>Limana, F., 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=Myocardial+infarction+induces+embryonic+reprogramming+of+epicardial+c-kit(+)+cells:+role+of+the+pericardial+fluid.">Myocardial infarction induces embryonic reprogramming of epicardial c-kit(+) cells: role of the pericardial fluid.</a> J Mol Cell Cardiol. 48 (4), 609-618 (2010).</li><li>Brunner, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+tissue+endothelin-1+levels+under+basal,+stimulated,+and+ischemic+conditions.">Cardiac tissue endothelin-1 levels under basal, stimulated, and ischemic conditions.</a> J Cardiovasc Pharmacol. 26, S44-S46 (1995).</li><li>de Lannoy, L. 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=Renin-angiotensin+system+components+in+the+interstitial+fluid+of+the+isolated+perfused+rat+heart.+Local+production+of+angiotensin+I.">Renin-angiotensin system components in the interstitial fluid of the isolated perfused rat heart. Local production of angiotensin I.</a> Hypertension. 29 (6), 1240-1251 (1997).</li><li>Strupp, M., Kammermeier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interstitial+Lactate+And+Glucose-Concentrations+Of+the+Isolated-Perfused+Rat-Heart+before,+during+And+after+Anoxia.">Interstitial Lactate And Glucose-Concentrations Of the Isolated-Perfused Rat-Heart before, during And after Anoxia.</a> Pflugers Arch. 423 (3-4), 232-237 (1993).</li><li>Wienen, W., Jungling, E., Kammermeier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enzyme-Release+into+the+Interstitial+Space+of+the+Isolated+Rat-Heart+Induced+by+Changes+in+Contractile+Performance.">Enzyme-Release into the Interstitial Space of the Isolated Rat-Heart Induced by Changes in Contractile Performance.</a> Cardiovasc Res. 28 (8), 1292-1298 (1994).</li><li>De Deckere, E. A., Ten Hoor, ., P, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+modified+Langendorff+technique+for+metabolic+investigations.">A modified Langendorff technique for metabolic investigations.</a> Pflugers Arch. 370 (1), 103-105 (1977).</li><li>Tschubar, F., Rose, H., Kammermeier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fatty+acid+transfer+across+the+myocardial+capillary+wall.">Fatty acid transfer across the myocardial capillary wall.</a> J Mol Cell Cardiol. 25 (4), 355-366 (1993).</li><li>Wienen, W., Kammermeier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-+and+extracellular+markers+in+interstitial+transudate+of+perfused+rat+hearts.">Intra- and extracellular markers in interstitial transudate of perfused rat hearts.</a> Am J Physiol. 254 (4 Pt 2), H785-H794 (1988).</li><li>Sasse, A., Ding, Z. P., Wallich, M., Godecke, A., Schrader, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+transfer+of+adenovirus+is+augmented+by+nitric+oxide+in+the+rat+heart.">Vascular transfer of adenovirus is augmented by nitric oxide in the rat heart.</a> Am J Physiol Heart Circ Physiol. 287 (3), H1362-H1368 (2004).</li><li>Gnecchi, M., Zhang, Z., Ni, A., Dzau, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paracrine+mechanisms+in+adult+stem+cell+signaling+and+therapy.">Paracrine mechanisms in adult stem cell signaling and therapy.</a> Circ Res. 103 (11), 1204-1219 (2008).</li><li>Herr, D. J., Aune, S. E., Menick, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+and+Assessment+of+Ischemia-reperfusion+Injury+in+Langendorff-perfused+Rat+Hearts.">Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts.</a> J Vis Exp. (101), e52908 (2015).</li><li>Ding, Z., 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=Epicardium-Derived+Cells+Formed+After+Myocardial+Injury+Display+Phagocytic+Activity+Permitting+In+Vivo+Labeling+and+Tracking.">Epicardium-Derived Cells Formed After Myocardial Injury Display Phagocytic Activity Permitting In Vivo Labeling and Tracking.</a> Stem Cells Transl Med. 5 (5), 639-650 (2016).</li><li>Hartwig, S., 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=Secretome+profiling+of+primary+human+skeletal+muscle+cells.">Secretome profiling of primary human skeletal muscle cells.</a> Biochim Biophys Acta. 1844 (5), 1011-1017 (2014).</li><li>Smolenski, R. T., Lachno, D. R., Ledingham, S. J. M., Yacoub, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+sixteen+nucleotides,+nucleosides+and+bases+using+high-performance+liquid+chromatography+and+its+application+to+the+study+of+purine+metabolism+in+hearts+for+transplantation.">Determination of sixteen nucleotides, nucleosides and bases using high-performance liquid chromatography and its application to the study of purine metabolism in hearts for transplantation.</a> J Chromatogr. 527 (2), 414-420 (1990).</li><li>Decking, U. K., Juengling, E., Kammermeier, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interstitial+transudate+concentration+of+adenosine+and+inosine+in+rat+and+guinea+pig+hearts.">Interstitial transudate concentration of adenosine and inosine in rat and guinea pig hearts.</a> Am J Physiol. 254 (6 Pt 2), H1125-H1132 (1988).</li><li>Heller, L. J., Mohrman, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimates+of+interstitial+adenosine+from+surface+exudates+of+isolated+rat+hearts.">Estimates of interstitial adenosine from surface exudates of isolated rat hearts.</a> J Mol Cell Cardiol. 20 (6), 509-523 (1988).</li></ol>

作者宣称他们没有竞争的经济利益。

逆心模型基于已建立的Langendorff心脏灌注技术12，并且通过简单地将心脏逆转到颠倒位置并使用刚性心室内球囊导管来保持该位置来执行。以这种方式，心脏间质渗出物可以与冠心静脉流出物灌注液物理分离，通过重力从心脏9的底部滴落。 CT可以通过放置在整个心脏表面上的薄而柔软的乳胶帽来连续收集。 该方法易于执行，除了Langendorff设备的成本最低。然而，一些步骤对于获得可重现和稳定的结果在技术上至关重要。这些包括确保乳胶帽适当地适合心脏的形状并且覆盖大约90％的心室表面而不是心房。时间从exc从动物的心脏到逆灌注的表现应该小于3分钟，因为长时间的缺血具有改变心脏代谢和形成心脏渗出的风险。通过心室内球囊测量的舒张性左心室压力应设定为填充心室（3-5 mmHg）。过度膨胀的气球可能会通过血管压迫改变冠状动脉血流。采样瓶（1.5 mL管）应保存在冰上，以避免感兴趣的代谢物和蛋白质的任何潜在降解。 此外，成功的实验在很大程度上取决于在心脏的准备，分离和插管期间的良好手动处理。这需要练习。为了保护心脏免受缺血性损伤，所有的准备工作都应该用冰冷的KHB进行。由于心脏的大小可能在大鼠之间变化，尽管体重相似，但建议使用轻微的适合不同尺寸的心脏的不同尺寸。 之前已经对孤立的大鼠5,6,7,8,9,10和豚鼠心脏16描述了分离的反向心脏方法，并且用于不同的目的。在本方法的描述中，我们对实验设置和样品处理进行了一些修改。例如，de Deckere 等人介绍的肺动脉插管如图7所示 ，这里没有进行，因为心脏的倒置位置可防止静脉流出物灌注液的潜在污染。通过降低CT放出管的开口引入轻微的负压，简化了CT的收集装置在反向心脏下方至10厘米。这使得更容易立即冷却渗出物样品。为了确保样品CT的快速冷却，通过将收集杯放入等体积的冰和NaCl混合物（ 即 1:1比例）中将收集杯预冷至-4℃的温度。这样可以快速冷却收集的CT样品。 通常，应该记住，分离的大鼠心脏与体内生理条件不同，因为间质液形成最可能小于盐水灌注心脏中的间质液形成。因此，分离心脏形成的CT可能不完全模拟体内组织液的真实组成。此外，本设置不能完全排除静脉流出物灌注液的潜在污染。然而，由于静脉出口位于心脏基底（直立心脏的最低水平），所以我们不相信这种污染有助于收集过程。 本方案描述了一种独特的方法，用于对心脏间质液进行采样，其中包含许多代谢物和蛋白质，其通过心肌细胞和非心脏细胞例如免疫细胞，内皮细胞，血管平滑肌细胞，成纤维细胞释放到间质液中，并且周细胞。心肌间质渗出物是通过内皮屏障10的流体输送以及一小部分淋巴液而形成的。它含有心脏代谢物7 ，间质空间中的可溶性因子和心脏和/或非心脏细胞的分泌物的混合物9 。因此，几种细胞类型有助于CT的形成。此外，还有几个影响形成速率的因素。首先，肿瘤压力似乎是调节反式激素的主要决定因素由于通过向灌注介质中加入葡聚糖或白蛋白使肿瘤血压升高显着降低CT 9,10的形成。其次，缺血9,16（包括心肌梗死）的血管通透性增加会增加灌注液的外渗，从而增加CT的形成。因此，在将来的研究中，肿瘤压力的增加可能是使CT体积最小化的合适手段，从而丰富了目标分子。

心衰（HF）是世界人类死亡的主要原因1 。 HF经常发生因心肌炎，心肌缺血损伤和左心室重塑，导致心脏收缩功能和患者生活质量逐渐恶化。虽然心脏病学和心脏手术的进步显着降低了HF死亡率，但它们仅仅是一个不可避免的进步性疾病过程的短暂的"延迟者"，具有显着的发病率。因此，目前缺乏有效的治疗强调需要鉴定可预防甚至逆转HF的新型分子靶标。这包括细胞外基质的改变，不受控的心脏免疫反应以及心脏和非心脏细胞之间的相互作用2 。 重要的是要认识到心脏细胞暴露于微环境的微环境tly塑造受伤心脏的免疫和再生反应。在分离的生理盐水灌注的心脏中，在生理和病理生理条件3,4,5下，以来自间质液空间（ 即微环境）的小液滴的形式在心脏表面产生CT。因此，CT（ 即组织间液）的分析可能有助于识别调节心脏代谢和收缩功能的因素6或影响移入受伤心脏后的免疫细胞功能。潜在地，这可能导致开发用于治疗HF的新型治疗策略。 从小鼠心脏收集CT在技术上是具有挑战性的。在常规Langendorff灌注的心脏中，CT的独家收集是困难的，因为CT与冠状动脉的混合静脉流出物灌注液不可预测地稀释从间质空间释放的任何浓度的代谢物/酶。克服这个限制的一个可能的策略是通过插入肺并同时连接肺静脉来排除静脉流出物7 。然而，这种方法面临与肺动脉和静脉的插管和结扎有关的困难，导致静脉流出物潜在渗漏到心脏渗出物中。使用反向心脏模型的概念首先由Kammermeier组引进，Kammermeier将其将分离的灌注心脏倒置到倒置位置，并将薄胶乳帽放置在心外膜表面上，以连续取样CT，而不会导致静脉流出物的污染8，9 。使用该程序，CT显示提供了从心脏9释放的代谢物的非常敏感的测量值，脂肪酸8和病毒颗粒10的毛细管转运。 最近，可能调节局部免疫应答和增加心脏血管生成的旁分泌因子11已经涉及干细胞治疗心脏病的有益作用。逆转心脏中CT的分析可能有助于化学鉴定这些个别旁分泌因子。此外，CT可能有助于确定心脏中免疫细胞体内活化所涉及的因素。 在这里提供的心脏表面CT收集的详细描述对于研究免疫细胞，成纤维细胞，内皮细胞和心肌细胞相对于整体心脏功能的相互作用的研究来说是实验性的。如上所述，间质液携带心脏内细胞间细胞通讯的信息，可以方便地通过收集CT来评估。详细的技术说明，包括如何从反向心脏收集CT的视频协议，应有助于今后应用这种独特的技术。

<table><tbody><tr><td>Latex Solution</td><td>ProChemie</td><td>Z-Latex LA-TZ</td><td>http://kautschukgesellschaft.de/%E2%80%A8z-latex-la-tz</td></tr><tr><td>Aluminum Mold</td><td>Home made</td><td>-</td><td>Reverse heart model</td></tr><tr><td>Universal Ovens</td><td>Memmert</td><td>UNB 400</td><td>Reverse heart model</td></tr><tr><td>Latex Balloon</td><td>Hugo Sachs</td><td>Size 4</td><td>Reverse heart model</td></tr><tr><td>Milling Machine</td><td>Proxxon</td><td>MF70</td><td>Reverse heart model</td></tr><tr><td>Sodium Chloride</td><td>Sigma</td><td>SZBD0810V</td><td>Chemicals</td></tr><tr><td>Sodium Hydrogen Carbonate</td><td>Roth</td><td>68852</td><td>Chemicals</td></tr><tr><td>Potassium Chloride</td><td>Merck</td><td>49361</td><td>Chemicals</td></tr><tr><td>Magnesium Sulphate Heptahydrate</td><td>Merck</td><td>58861</td><td>Chemicals</td></tr><tr><td>Potassium Dihydrogen Phosphate</td><td>Merck</td><td>48731</td><td>Chemicals</td></tr><tr><td>D(+)-Glucose Anhydrous</td><td>Merck</td><td>83371</td><td>Chemicals</td></tr><tr><td>Calcium Chloride Dihydrate</td><td>Fluka</td><td>21097</td><td>Chemicals</td></tr><tr><td>Balance</td><td>VWR</td><td>SE 1202</td><td>Weighing chemicals</td></tr><tr><td>Double Distilled Water</td><td>Millpore</td><td>-</td><td>Disolving chemicals</td></tr><tr><td>Medical Pressure Transducer</td><td>Gold</td><td>-</td><td>Langendorff apparatus</td></tr><tr><td>Medical Flow Probe</td><td>Transonic</td><td>3PXN</td><td>Langendorff apparatus</td></tr><tr><td>Heating Circulating Bath</td><td>Haake&amp;nbsp;</td><td>B3 ; DC1</td><td>Langendorff apparatus</td></tr><tr><td>Laboratory and Vaccum Tubing</td><td>Tygon</td><td>R-3603</td><td>Langendorff apparatus</td></tr><tr><td>Animal Research Flowmeters</td><td>Transonic</td><td>T206</td><td>Langendorff apparatus</td></tr><tr><td>PowerLab Data Acquisition Device</td><td>AD Instruments</td><td>Chart 7.1</td><td>Langendorff apparatus</td></tr><tr><td>LabChart Data Acquisition Software</td><td>AD Instruments</td><td>Chart 7.1</td><td>Langendorff apparatus</td></tr><tr><td>Peristaltic Pump</td><td>Glison</td><td>MINIPULS 3</td><td>Langendorff apparatus</td></tr><tr><td>Glass Water Column</td><td>home made</td><td>-</td><td>Langendorff apparatus</td></tr><tr><td>Water Bath Protective Agent</td><td>VWR</td><td>462-7000</td><td>Langendorff apparatus</td></tr><tr><td>Sterile Disposable Filters (0.2 &amp;micro;m)</td><td>Thermo Scientific</td><td>595-4520</td><td>Langendorff apparatus</td></tr><tr><td>Blood gas analyzers</td><td>Radiometer</td><td>ABL90 FLEX PLUS</td><td>Gas analyzer</td></tr><tr><td>70% ethanol</td><td>VWR</td><td>UN1170</td><td>Cleaning&amp;nbsp; tubings</td></tr><tr><td>100% ethanol</td><td>Merck</td><td>64-17-5</td><td>Cleaning tubings</td></tr><tr><td>Wistar Rats</td><td>Janvier</td><td>-</td><td>Animals</td></tr><tr><td>Stainless Scissors</td><td>AESCULAP</td><td>BC702R</td><td>Surgical Instruments</td></tr><tr><td>Stainless Scissors</td><td>AESCULAP</td><td>BC257R</td><td>Surgical Instruments</td></tr><tr><td>Big Forceps</td><td>AESCULAP</td><td>-</td><td>Surgical Instruments</td></tr><tr><td>8m/m Stainless Forceps</td><td>F.S.T</td><td>11052-10</td><td>Surgical Instruments</td></tr><tr><td>superfine (10/0) emery paper</td><td>3M</td><td>051111-11694</td><td>Reverse heart model</td></tr></tbody></table>

the inverted heart model for interstitial transudate collection from the isolated rat heart

本协议描述了一种独特的方法，可以从分离的盐水灌注的大鼠心脏中收集心脏渗出物（CT）。根据Langendorff技术分离和逆行灌注心脏后，将心脏倒置为倒立位置，并通过插入左心室的气囊导管进行机械稳定。然后，预先浇铸以匹配大鼠心脏的平均尺寸的薄胶乳盖被放置在心外膜表面上。胶乳盖的出口连接到硅管，远端开口比心脏底部高10厘米，产生轻微的吸力。在心外膜表面连续生产的CT被收集在冰冷的小瓶中用于进一步分析。对照组和梗死心脏CT值分别为17〜147μL/ min（n = 14），占冠状静脉血液灌注液0.1-1％。蛋白质组学分析和高度信息rmance液相色谱（HPLC）显示收集的CT含有广谱的蛋白质和嘌呤能代谢物。

<html>逆心模型能够在分离的，逆灌注的大鼠心脏中收集心脏间质渗出物（ 图1A- C ）。当在100cmH 2 O的恒定压力下灌注时，组织液形成的速率在17和147μL/ min之间，占分离心脏中冠状静脉流出物的0.1-1％。 用二喹啉酸（BCA）测定的CT的蛋白质含量为1.08±0.40mg / mL（n = 6）。一维凝胶电泳分析（SDS-PAGE）显示了心脏渗出液中存在的广谱蛋白质（ 图2A ）。在进行50分钟缺血/再灌注的心脏上进行二维荧光差异凝胶电泳（2D-DIGE）。如图所示<html>图2B中，发现缺血性心脏CT中有几种蛋白质上调，其中70.1％为细胞外基质蛋白，4.6％为细胞膜蛋白，17.2％为细胞质蛋白，2.3％为核蛋白质（ 表1 ）。 嘌呤一直被认为是调节心脏免疫反应，血管舒缩功能和心脏功能的关键信号分子，特别是缺血性损伤后。 CT的收集允许在病理生理条件如MI下测量存在于心脏间质液中的各种代谢物。 如图3所示，通过HPLC测量的AMP，GMP，NADP，腺苷，次黄嘌呤和尿酸的浓度在缺血性心脏中较高，这与先前使用其它方法16报道的结果相似，class ="xref"&gt; 17。 <img alt="表格1" src="/files/ftp_upload/55849/55849table1.jpg" /> 表1:缺血性心脏CT的上调蛋白质列表。通过2D-DIGE分析来自缺血性心脏的CT，并通过蛋白质组学鉴定。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55849/55849_Table_1_protein_IDs.xls">请点击这里下载此表。</a> <img alt="图1" class="xfigimg" src="/files/ftp_upload/55849/55849fig1.jpg" /> 图1:逆心模型的示意图。 （ A ）由适当形状和尺寸的大鼠心脏构建铝模具。 （ B ）将模具浸渍到胶乳溶液中后，铸造厚度约0.01mm的胶乳盖。 （ C ）在L心脏放射心脏设备，使用主动脉插管通过主动脉灌注心脏，后者将其重新倒置成倒立位置，并由置于左心室的心室内球囊机械支撑。通过压力传感器监测室内压力的发展。乳胶帽覆盖了右心室和左心室表面的近90％，出口连接到硅管（ID = 0.2mm），远端开口距心脏底部10cm。这产生轻微负的静水压力。心脏渗出液 通常被收集到冰冷却的1.5mL管中。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55849/55849fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图2" class="xfigimg" src="/files/ftp_upload/55849/55849fig2.jpg" /> 图2:CT的整体分析。通过SDS-PAGE（ A ）分离CT中的蛋白质，并通过2D-DIGE分析（ B ）鉴定。对于（A），泳道1-4表示来自个体心脏的心脏样品（1和2 =假手术; 3和4 =梗塞）。对于（B），在梗死心脏的CT上进行2D-DIGE。通过液相色谱（LC）-MS / MS 14,15证实蛋白质同一性。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55849/55849fig2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图3" class="xfigimg" src="/files/ftp_upload/55849/55849fig3.jpg" /> 图3:CT中的嘌呤。通过HPLC分析存在于CT中的各种嘌呤。代表性的HPLC从假手术（蓝色）和梗死（黑色）心脏显示，梗死心脏呈现较高的AMP和腺苷的间质浓度，而不是次黄嘌呤。 <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55849/55849fig3large.jpg" target="_blank">请点击此处查看此图的较大版本。</a>

Watch this Scientific Journal Video about The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart at JoVE.com

The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart

该方案描述了从分离的灌注大鼠心脏中收集心脏间质液的方法。为了将间质渗出物与冠状静脉流出物灌注液物理分离，Langendorff灌注的心脏被倒置，并且使用软胶乳帽收集在心脏表面上形成的渗出物（间质液体）。

从孤立的大鼠心脏的间质渗出物收集的倒置心脏模型

The present protocol describes a unique approach that enables the collection of cardiac transudate (CT) from the isolated, saline-perfused rat heart. ...

the-inverted-heart-model-for-interstitial-transudate-collection-from

Universidad San Sebastian

Research

JoVE Journal

Medicine

8.3K 次观看.  Second Military Medical University. 该方案描述了从分离的灌注大鼠心脏中收集心脏间质液的方法。为了将间质渗出物与冠状静脉流出物灌注液物理分离，Langendorff灌注的心脏被倒置，并且使用软胶乳帽收集在心脏表面上形成的渗出物（间质液体）。 

视频: The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart

An Isolated Working Heart System for Large Animal Models

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart model, have been invaluable tools for studying cardiovascular function and disease1-15. Although the Langendorff heart preparation can be used for any mammalian heart, most studies involving this apparatus use small animal models (e.g., mouse, rat, and rabbit) due to the increased complexity of systems for larger mammals1,3,11. One major difficulty is ensuring a constant coronary perfusion pressure over a range of different heart sizes &#8211; a key component of any experiment utilizing this device1,11. By replacing the classic hydrostatic afterload column with a centrifugal pump, the Langendorff working heart apparatus described below allows for easy adjustment and tight regulation of perfusion pressures, meaning the same set-up can be used for various species or heart sizes. Furthermore, this configuration can also seamlessly switch between constant pressure or constant flow during reperfusion, depending on the user&#8217;s preferences. The open nature of this setup, despite making temperature regulation more difficult than other designs, allows for easy collection of effluent and ventricular pressure-volume data.

Most studies involving the Langendorff apparatus use small animal models due to the increased complexity of systems for larger mammals. We describe a Langendorff system for large animal models that allows for use across a range of species, including humans, and relatively easy data acquisition.

一个孤立的工作心脏系统的大型动物模型

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart ...

科研

医学

Isolation of Endocardial and Coronary Endothelial Cells from the Ventricular Free Wall of the Rat Heart

It has been shown that endocardial endothelial cells (EECs) and coronary endothelial cells (CECs) differ in origin, development, markers, and functions. Consequently, these two cell populations play unique roles in cardiac diseases. Current studies involving isolated endothelial cells investigate cell populations consisting of both EECs and CECs. This protocol outlines a method to independently isolate these two cell populations for cell-specific characterization. Following the collection of the left and right ventricular free wall, endothelial cells from the outer surface and inner surface are separately liberated using a digestion buffer solution. The sequential digestion of the outer surface and the inner endocardial layer retained separation of the two endothelial cell populations. The separate isolation of EECs and CECs is further verified through the identification of markers specific to each population. Based on previously published single cell RNA profiling in the mouse heart, the Npr3, Hapln1, and Cdh11 gene expression is unique to EECs; while Fabp4, Mgll, and Cd36 gene expression is unique to CECs. qPCR data revealed enriched expression of these characteristic markers in their respective samples, indicating successful EEC and CEC isolation, as well as maintenance of cell phenotype, enabling further cell-specific functional analysis.

We present a protocol for the isolation of endocardial and coronary endothelial cells from rat hearts through sequential tissue digestion in a digestion buffer, cell collection from recurrent centrifuge cycles, and cell purification using anti-rat CD31 microbeads.

从鼠心心室自由壁分离内膜细胞和冠状内皮细胞

It has been shown that endocardial endothelial cells (EECs) and coronary endothelial cells (CECs) differ in origin, development, markers, and functions. ...

发育生物学

An Immunological Model for Heterotopic Heart and Cardiac Muscle Cell Transplantation in Rats

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications have been reported since the first description in 1964. After 30 years of performing heterotopic heart transplantation in rats, we have developed a simplified surgical approach, which can be easily taught and performed without further surgical training or background.
After dissection of the ascending aorta and the pulmonary artery and ligation of superior and inferior caval and pulmonary veins, the donor heart is harvested and subsequently perfused with ice-cold saline solution supplemented with heparin. After clamping and incising the recipient abdominal vessels, the donor ascending aorta and pulmonary artery are anastomosed to the recipient abdominal aorta and inferior vena cava, respectively, using continuous running sutures.
Depending on different donor-recipient combinations, this model allows analyses of either acute or chronic rejection of allografts. The immunological significance of this model is further enhanced by a novel approach of in-ear injection of vital cardiac muscle cells and subsequent analysis of draining cervical lymphatic tissue.

We describe a model of heterotopic abdominal heart transplantation in rats, implying modifications of current strategies, which lead to a simplified surgical approach. Additionally, we describe a novel rejection model by in-ear injection of vital cardiac muscle cells, allowing further transplant immunological analyses in rats.

大鼠异位心脏和心肌细胞移植的免疫模型

Heterotopic heart transplantation in rats has been a commonly used model for diverse immunological studies for more than 50 years. Several modifications ...

免疫与感染

Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of metabolic syndrome (MetS)-related heart failure with preserved ejection fraction (HFpEF). The prevalence of MetS-related HFpEF is rising, and atrial cardiomyopathies associated with atrial remodeling and atrial fibrillation are clinically highly relevant as atrial remodeling is an independent predictor of mortality. Studies with isolated single-cell cardiomyocytes are frequently used to corroborate and complement in vivo findings. Circulatory vessel rarefication and interstitial tissue fibrosis pose a potentially limiting factor for the successful single-cell isolation of ACMs from animal models of this disease.
We have addressed this issue by employing a device capable of manually regulating the intraluminal pressure of cardiac cavities during the isolation procedure, substantially increasing the yield of morphologically and functionally intact ACMs. The acquired cells can be used in a variety of different experiments, such as cell culture and functional Calcium imaging (i.e., excitation-contraction-coupling).
We provide the researcher with a step-by-step protocol, a list of optimized solutions, thorough instructions to prepare the necessary equipment, and a comprehensive troubleshooting guide. While the initial implementation of the procedure might be rather difficult, a successful adaptation will allow the reader to perform state-of-the-art ACM isolations in a rat model of MetS-related HFpEF for a broad spectrum of experiments.

Here, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes from a rat model of metabolic syndrome-related heart failure with preserved ejection fraction. A manual regulation of intraluminal pressure of cardiac cavities is implemented to yield functionally intact myocytes suitable for excitation-contraction-coupling studies.

代谢综合征心力衰竭模型大鼠心房心肌细胞的分离与保留射血分数的关系

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of ...

A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of transplant with the potential to deliver durable heart valves capable of somatic growth with no requirement for anticoagulation. However, the immunobiology of heart valve transplants remains unexplored, highlighting the need for animal models to study this new type of transplant. Previous rat models for heterotopic aortic valve transplantation into the abdominal aorta have been described, though they are technically challenging and costly. For addressing this challenge, a renal subcapsular transplant model was developed in rodents as a practical and more straightforward method for studying heart valve transplant immunobiology. In this model, a single aortic valve leaflet is harvested and inserted into the renal subcapsular space. The kidney is easily accessible, and the transplanted tissue is securely contained in a subcapsular space that is well vascularized and can accommodate a variety of tissue sizes. Furthermore, because a single rat can provide three donor aortic leaflets and a single kidney can provide multiple sites for transplanted tissue, fewer rats are required for a given study. Here, the transplantation technique is described, providing a significant step forward in studying the transplant immunology of heart valve transplantation.

This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.

啮齿动物异位心脏瓣膜移植的简化模型

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of ...

Rodent Working Heart Model for the Study of Myocardial Performance and Oxygen Consumption

Isolated working heart models have been used to understand the effects of loading conditions, heart rate and medications on myocardial performance in ways that cannot be accomplished in vivo. For example, inotropic medications commonly also affect preload and afterload, precluding load-independent assessments of their myocardial effects in vivo. Additionally, this model allows for sampling of coronary sinus effluent without contamination from systemic venous return, permitting assessment of myocardial oxygen consumption. Further, the advent of miniaturized pressure-volume catheters has allowed for the precise quantification of markers of both systolic and diastolic performance. We describe a model in which the left ventricle can be studied while performing both volume and pressure work under controlled conditions. 
In this technique, the heart and lungs of a Sprague-Dawley rat (weight 300-500 g) are removed en bloc under general anesthesia. The aorta is dissected free and cannulated for retrograde perfusion with oxygenated Krebs buffer. The pulmonary arteries and veins are ligated and the lungs removed from the preparation. The left atrium is then incised and cannulated using a separate venous cannula, attached to a preload block. Once this is determined to be leak-free, the left heart is loaded and retrograde perfusion stopped, creating the working heart model. The pulmonary artery is incised and cannulated for collection of coronary effluent and determination of myocardial oxygen consumption. A pressure-volume catheter is placed into the left ventricle either retrograde or through apical puncture. If desired, atrial pacing wires can be placed for more precise control of heart rate. This model allows for precise control of preload (using a left atrial pressure block), afterload (using an afterload block), heart rate (using pacing wires) and oxygen tension (using oxygen mixtures within the perfusate).

Isolated working heart models can be used to measure the effect of loading conditions, heart rate, and medications on myocardial performance and oxygen consumption. We describe methods for preparation of a rodent left heart working model that permits study of systolic and diastolic performance and oxygen consumption under various conditions.

灭鼠工作心脏模型心肌的性能和氧耗的研究

Isolated working heart models have been used to understand the effects of loading conditions, heart rate and medications on myocardial performance in ways ...

生物学

A Heterotopic Rat Heart Transplantation Model using Circulatory Death Donor Hearts

The objective of this protocol is to set up a rat heterotopic heart transplantation model with donation after circulatory death (DCD) donor hearts. There are two setups for this protocol: heart donor setup and recipient setup. In the heart donor setup, Sprague Dawley rats are anesthetized, endotracheally intubated, and ventilated. The right carotid artery is cannulated to deliver heparin and the paralytic agent vecuronium-bromide. The DCD process is initiated by terminating the ventilation. After 20 min, the heart is exposed and the aorta distal to the brachiocephalic branch is clamped. At 25 min from terminating the ventilator, ice-cold University of Wisconsin (UW) solution is perfused through the carotid catheter to flush the heart. The heart is procured by dividing the aorta, pulmonary artery, venae cavae, and pulmonary veins and stored in UW solution for implantation. In the recipient setup, the Lewis rat is anesthetized with isoflurane. Slow-release buprenorphine is administered subcutaneously to facilitate a smooth postoperative recovery. Through a midline abdominal incision, the infra-renal aorta and the inferior vena cava are isolated and clamped with an atraumatic vascular clamp. The donor heart aorta and pulmonary artery are sutured to the recipient abdominal aorta and vena cava, respectively, with a running 8-0 Prolene. The vascular clamp is removed to reperfuse the heart. The abdominal wall is closed and the rat is recovered. After a set interval (24 h to 2 weeks), the recipient rat is anesthetized, the transplanted heart is exposed, and a balloon-tip-catheter is inserted into the left ventricle via the apex to record developed pressure and dP/dt using a data acquisition system. The heart tissue is collected for histology, immunology, or molecular analysis. A successful DCD donor rat heart transplantation model will allow further studies on the cardioprotective approaches to improve heart transplantation outcomes from DCD donors.

Here, we describe the preparation and technical details of murine heterotopic heart transplantation utilizing a circulatory death donor heart.

The objective of this protocol is to set up a rat heterotopic heart transplantation model with donation after circulatory death (DCD) donor hearts. There ...

A Model of Long-Term Ventricular Fibrillation in Isolated Rat Hearts

Ventricular fibrillation (VF) is a fatal arrhythmia with a high incidence in cardiac patients, but VF arrest under perfusion is a neglected method of intraoperative arrest in the field of cardiac surgery. With recent advances in cardiac surgery, the demand for prolonged VF studies under perfusion has increased. However, the field lacks simple, reliable, and reproducible animal models of chronic ventricular fibrillation. This protocol induces long-term VF through alternating current (AC) electrical stimulation of the epicardium. Different conditions were used to induce VF, including continuous stimulation with a low or high voltage to induce long-term VF and stimulation for 5 min with a low or high voltage to induce spontaneous long-term VF. The success rates of the different conditions, as well as the rates of myocardial injury and recovery of cardiac function, were compared. The results showed that continuous low-voltage stimulation induced long-term VF and that 5 min of low-voltage stimulation induced spontaneous long-term VF with mild myocardial injury and a high rate of recovery of cardiac function. However, the low-voltage, continuously stimulated long-term VF model had a higher success rate. High-voltage stimulation provided a higher rate of VF induction but showed a low defibrillation success rate, poor recovery of cardiac function, and severe myocardial injury. On the basis of these results, continuous low-voltage epicardial AC stimulation is recommended for its high success rate, stability, reliability, reproducibility, low impact on cardiac function, and mild myocardial injury.

This protocol presents a model of long-term ventricular fibrillation in rat hearts induced by continuous stimulation with low-voltage alternating current. This model has a high success rate, is stable, reliable, and reproducible, has a low impact on cardiac function, and causes only mild myocardial injury.

孤立大鼠心脏长期心室颤动模型

Ventricular fibrillation (VF) is a fatal arrhythmia with a high incidence in cardiac patients, but VF arrest under perfusion is a neglected method of ...

Rat Model of Normothermic Ex-Situ Perfused Heterotopic Heart Transplantation

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the number of heart failure patients waiting for transplantation is still increasing. The normothermic ex situ preservation technique has been established as a comparable method to the conventional static cold storage technique. The main advantage of this technique is that donor hearts can be preserved for up to 12 h in a physiologic condition. Moreover, this technique allows resuscitation of the donor hearts after circulatory death and applies required pharmacologic interventions to improve donor function after implantation. Numerous animal models have been established to improve normothermic ex situ preservation techniques and eliminate preservation-related complications. Although large animal models are easy to handle compared to small animal models, it is costly and challenging. We present a rat model of normothermic ex situ donor heart preservation followed by heterotopic abdominal transplantation. This model is relatively cheap and can be accomplished by a single experimenter.

Here, we present an assessment protocol of a heterotopically implanted heart after normothermic ex situ preservation in the rat model.

异位灌注异位心脏移植大鼠模型

Heart transplantation is the most effective therapy for end-stage heart failure. Despite the improvements in therapeutic approaches and interventions, the ...

Modified Heterotopic Abdominal Heart Transplantation and a Novel Aortic Regurgitation Model in Rats

Over the past 50 years, many researchers have reported heterotopic abdominal heart transplantation in mice and rats, with some variations in the surgical technique. Modifying the transplantation procedure to strengthen the myocardial protection could prolong the ischemia time while preserving the donor&#39;s cardiac function. This technique&#39;s key points are as follows: transecting the donor&#39;s abdominal aorta before harvesting to unload the donor&#39;s heart; perfusing the donor&#39;s coronary arteries with a cold cardioplegic solution; and topical cooling of the donor&#39;s heart during the anastomosis procedure. Consequently, since this procedure prolongs the acceptable ischemia time, beginners can easily perform it and achieve a high success rate.
Moreover, a new aortic regurgitation (AR) model was established in this work using a technique different from the existing one,&#160;which is created by inserting a catheter from the right carotid artery and puncturing the native aortic valve under continuous echocardiographic guidance. A heterotopic abdominal heart transplantation was performed using the novel AR model. In the protocol, after the donor&#39;s heart is harvested, a stiff guidewire is inserted into the donor&#39;s brachiocephalic artery and advanced toward the aortic root. The aortic valve is punctured by pushing the guidewire further even after the resistance is felt, thus inducing AR. It is easier to damage the aortic valve using this method than with the procedure described in the conventional AR model. Additionally, this novel AR model does not contribute to the recipient&#39;s circulation; therefore, this method is expected to produce a more severe AR model than the conventional procedure.

This study demonstrates a reproducible heterotopic abdominal heart transplantation technique in rats that beginners can learn and perform. Additionally, a novel aortic regurgitation model in rats is generated by performing heterotopic abdominal heart transplantation and damaging the donor's aortic valve using a guidewire after harvesting.

改良异位腹心移植及新型大鼠主动脉瓣反流模型

Over the past 50 years, many researchers have reported heterotopic abdominal heart transplantation in mice and rats, with some variations in the surgical ...