这项工作是从SNF [31003A-138528]，该Octav和马塞拉Botnar基金会，诺华基金会和艾玛Muschamp基金会的资助。我们感谢马丁Lambelet，和让 - 克里斯托夫施特勒的出色技术援助。

洛桑大学的伦理委员会批准了试验，这是按照1975年赫尔辛基宣言提出的原则，修订于1983年为使用人体组织。 1，人类大隐静脉丰收<ol><li>从发生下肢搭桥手术缺血患者获得非曲张的大隐静脉的盈余部分。在手术室，消毒与碘溶液的整个腿和悬垂性，患者从腹股沟到脚露出的腿。 </li><li>请从腹股沟到膝盖正中切口（在离开中断的皮肤部分）。 </li><li>收获大隐静脉与周围组织（非接触式技术）的蒂。用4-0丝束静脉安全侧枝。在4℃下立即存储最小为9厘米的大隐静脉的长剩余段，具有2.5-4毫米的外径在RPMI-1640ģlutamax介质，辅以12.5％胎牛血清，并把它带到实验室。 </li></ol> 2，每股内含价值的设计<ol><li>装配在图1所示的通用设备。高压灭菌的所有设备，并保持所有组件在无菌条件下。此外，确保系统具有防水和不漏化学品到培养基中。为封面用聚甲基丙烯酸甲酯（PMMA-GS）。钢（X5铬镍18 10）和聚甲醛塑料（聚甲醛）作为静脉的支持。 </li><li>设计灌注腔至所需的几何形状，以使放置在静脉和其连接的。确保深度（或半径如果使用圆柱形结构）是至少2.5厘米所以它允许随着恒定覆盖的培养基（ 图1）最小屈曲容器和扩张。密封是一个主要问题，是长方形的PMMA-GS建设使用的原因。 </li><li>设计静脉支持所需的geometr年。避免静脉扭结或以上腹胀，允许长度调节通过推动或拉动（螺钉不能用于这一目的，因为静脉将与螺钉扭沿）。 注:完全钢杆被2连接滑动的L形件支持2静脉缸（直径5毫米，以适应容器）和静脉（ 图1B和图2）被用在这里。 </li><li>设计压力柱，使得施加到该系统的"静息压力"是:P = 0-10 = HXρXG，其p =压力（N / m 2时，Pa）的流体柱（米）为h =高度ρ =液体（千克/米3）和g =重力常数（9.81米/秒2）的密度。设计四个连接管，从顶部到底部:以施加压力，对所述流出（从静脉），所述流入（到静脉），并允许更换培养基。 </li><li>准备中。根据以往的研究5,11-14，选择的RPMI-1640，辅以Glutamax的，12.5％胎牛血清和1％抗生素 - 抗真菌剂溶液（10,000 U / ml的青霉素G，加上10毫克/毫升硫酸链霉素，外加25毫克/毫升两性霉素B，加0.5微克/毫升:庆大霉素）。剪切应力（SS）由SS = 4μQ/π* R 3 Q给定的流速（毫升/秒），R的静脉段的半径（厘米），并且μ是灌注介质的粘度。 <ol><li>调制的SS通过另外的70 kDa的葡聚糖调节粘度。测量用粘度计的粘度。这里，加8％70kDa的葡聚糖设置SS到9-15达因/厘米2。 </li></ol></li><li>设置齿轮泵诱导的60个脉冲/分钟，恒定幅度产生的150±15毫升/分钟的单向流动，独立于由计算机系统中的应用和控制的压力脉动心形信号。确保该驱动软件集成恒定采集与监控压力，流速，脉搏速率，和信号。如果需要，使用第二个泵（非同步hronized），以产生一个非层流，湍流。 </li></ol> 3 EVPS组件（图1） <ol><li>在开始之前，请确保所有的设备是无菌的。执行层流罩下无菌以下所有步骤。 </li><li>将静脉在培养皿中充满介质。使用手术刀片，分化静脉分成3等份。 </li><li>立即冲洗的PBS一段。划分段三部分，在福尔马林固定一个用于形态。冻结的其它两个用于定量转录物（RT-PCR）和蛋白质（免疫印迹）分析。考虑这些片段作为对照，非静脉灌注。 </li><li>用剩下的2段灌注。 <ol><li>轻轻地注入介质插入静脉并确定正常流动方向;在阀门存在的静脉是相反的。 </li><li>密封脉是最重要的实验取得成功。检查通过络脉泄漏。争取用6-0丝线缝合任何泄漏。 </li> &lt;/ OL&gt; </li><li>连接这两个金属圆柱体之间的静脉段，一端在一个时间（2.3， 图1）。确保与Ethibon 3-0气缸周围的凹部（ 图1A和B）。 <ol><li>将整个静脉段进入灌注腔预先填充有介质。重复同样的步骤进行的第二部分。 注意:不正确密封静脉缸将泄漏的来源，需要再次介入，并显著增加感染和实验失败的风险。 </li></ol></li><li>加强（目）的第二段，松开两个汽缸（装有静脉）从L形件（2.3和图1）。 <ol><li>要轻柔，不要触碰静脉使用任何手段。第一张幻灯片网格上的圆筒，然后到静脉。推/拉推撞就会在静脉网。 </li><li>一旦网格覆盖静脉的整个表面上固定的第jacketed静脉与Ethibon 3-0缸。 </li><li>重新组装静脉/气缸化合物为L形支撑，并将其转移到灌注腔，预先填充有介质。 </li></ol></li><li>使用过氧化处理的硅橡胶管具有3.2毫米的内径（在和流出），每个金属筒连接到一个Y型分离器。 </li><li>连接的流出分离器使用相同类型管道的第二Y型分离器。从这个Y型分线器，​​使用一管来测量通过两艘船的灌注压。连接的另一个回柱，以形成一个封闭的回路系统（ 图2）。 </li><li>内的培养箱中，使用长（一米长）管与低压塔连接到泵头部。 </li><li>完成建立连接的泵头部，以在流入Y型分光器与另一个长长度管（ 图1）。 </li></ol> 4，静脉灌注<ol><li>之后的每股内含价值组件已经合作mpleted，填充介质柱（保持低于静脉流出道，让加气站）。添加更多的介质进入塔之前，系统已满。将所有系统进入维持在37±0.1℃，pH值为孵化器保持恒定在7.40±0.01（使用基于恒基哈塞尔巴赫方程的CO 2 / pH值算法）。 </li><li>使齿轮泵头部培养箱外，并将其连接到齿轮泵的驱动。螺丝杆，以确保大会。 </li><li>交换机上的泵浦功率，确保其在驱动软件激活，允许5分钟的灭火剂能均匀地分布在每一个车厢。 </li><li>要监视的压力，用动脉在线监测。连接EVPS压力输出（它对应于动脉导管），以连接到计算机中的压力换能器。 <ol><li>确保将管完全充满介质，并且不包含任何气泡。脱泡的培养体系，通过"动脉里NE"管（ 图2）。注意显示器，寻找60个脉冲/恒定振幅的分的心搏动信号。在这点上，平均压力为0-10毫米汞柱之间。如果压力&lt;0且列逐渐清空寻找（静脉和管之间的静脉侧支或密封不足）的泄漏。 </li></ol></li><li>设置最小压力至6毫米汞柱的静脉试验或在90毫米汞柱为动脉的测试。在这些条件下，空气喷射器施加所需压力的柱和系统。 </li><li>通过使用连接到高压塔的管更换培养基，每2天。为了防止压力变化造成的破坏，先打开柱插头。 </li></ol> 5，完成灌注的<ol><li>灌注后3天或7天:乘坐每股内含价值从孵化器和下马的静脉。丢弃5毫米近端和远端静脉端连接于设备。切中央5毫米厚的RIN从剩余的部分GS和福尔马林（形态）修复。冻结剩余的碎片和降低成粉末作进一步的分子分析。 </li></ol>

<ol>
	<li>Sal Go, A., 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=Executive+summary:+heart+disease+and+stroke+statistics--2014+update:+a+report+from+the+american+heart+association.">Executive summary: heart disease and stroke statistics--2014 update: a report from the american heart association.</a> Circulation. 129, 399-410 (2014).</li><li>Sal Conte, 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=Results+of+PREVENT+III:+a+multicenter,+randomized+trial+of+edifoligide+for+the+prevention+of+vein+graft+failure+in+lower+extremity+bypass+surgery.">Results of PREVENT III: a multicenter, randomized trial of edifoligide for the prevention of vein graft failure in lower extremity bypass surgery.</a> Journal of Vascular Surgery. 43, 742-751 (2006).</li><li>Yu, P., Nguyen, B. T., Tao, M., Bai, Y., Ozaki, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+vein+graft+hemodynamic+manipulations+to+enhance+experimental+utility.">Mouse vein graft hemodynamic manipulations to enhance experimental utility.</a> The American Journal of Pathology. 178, 2910-2919 (2011).</li><li>Davies, M. G., Hagen, P. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reprinted+article+&amp;#34;Pathophysiology+of+vein+graft+failure:+a+review&amp;#34;.">Reprinted article &amp;#34;Pathophysiology of vein graft failure: a review&amp;#34;.</a> European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery. 42, S19-S29 (2011).</li><li>Berard, X., 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=Role+of+hemodynamic+forces+in+the+ex+vivo+arterialization+of+human+saphenous+veins.">Role of hemodynamic forces in the ex vivo arterialization of human saphenous veins.</a> Journal of Vascular Surgery. 57, 1371-1382 (2013).</li><li>Vijayan, V., 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=Long-term+reduction+of+medial+and+intimal+thickening+in+porcine+saphenous+vein+grafts+with+a+polyglactin+biodegradable+external+sheath.">Long-term reduction of medial and intimal thickening in porcine saphenous vein grafts with a polyglactin biodegradable external sheath.</a> Journal of Vascular Surgery. 40, 1011-1019 (2004).</li><li>Jeremy, J. Y., 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=On+the+biology+of+saphenous+vein+grafts+fitted+with+external+synthetic+sheaths+and+stents.">On the biology of saphenous vein grafts fitted with external synthetic sheaths and stents.</a> Biomaterials. 28, 895-908 (2007).</li><li>Zilla, P., 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=Constrictive+external+nitinol+meshes+inhibit+vein+graft+intimal+hyperplasia+in+nonhuman+primates.">Constrictive external nitinol meshes inhibit vein graft intimal hyperplasia in nonhuman primates.</a> The Journal of Thoracic and Cardiovascular Surgery. 136, 717-725 (2008).</li><li>Zilla, P., 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=Utilization+of+shape+memory+in+external+vein-graft+meshes+allows+extreme+diameter+constriction+for+suppressing+intimal+hyperplasia:+a+non-human+primate+study.">Utilization of shape memory in external vein-graft meshes allows extreme diameter constriction for suppressing intimal hyperplasia: a non-human primate study.</a> Journal of Vascular Surgery. 49, 1532-1542 (2009).</li><li>Yeoman, M. 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=A+constitutive+model+for+the+warp-weft+coupled+non-linear+behavior+of+knitted+biomedical+textiles.">A constitutive model for the warp-weft coupled non-linear behavior of knitted biomedical textiles.</a> Biomaterials. 31, 8484-8493 (2010).</li><li>Longchamp, A., 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=The+use+of+external+mesh+reinforcement+to+reduce+intimal+hyperplasia+and+preserve+the+structure+of+human+saphenous+veins.">The use of external mesh reinforcement to reduce intimal hyperplasia and preserve the structure of human saphenous veins.</a> Biomaterials. 35, 2588-2599 (2014).</li><li>Saucy, 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=Ex+vivo+pulsatile+perfusion+of+human+saphenous+veins+induces+intimal+hyperplasia+and+increased+levels+of+the+plasminogen+activator+inhibitor+1.">Ex vivo pulsatile perfusion of human saphenous veins induces intimal hyperplasia and increased levels of the plasminogen activator inhibitor 1.</a> European Surgical Research. Europaische Chirurgische Forschung. Recherches Chirurgicales Europeennes. 45, 50-59 (2010).</li><li>Dubuis, C., 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=Atorvastatin-loaded+hydrogel+affects+the+smooth+muscle+cells+of+human+veins.">Atorvastatin-loaded hydrogel affects the smooth muscle cells of human veins.</a> The Journal of pharmacology and experimental. 347, 574-581 (2013).</li><li>Deglise, 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=Increased+connexin43+expression+in+human+saphenous+veins+in+culture+is+associated+with+intimal+hyperplasia.">Increased connexin43 expression in human saphenous veins in culture is associated with intimal hyperplasia.</a> Journal of Vascular Surgery. 41, 1043-1052 (2005).</li><li>Muto, A., Model, L., Ziegler, K., Eghbalieh, S. D., Dardik, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+vein+graft+adaptation+to+the+arterial+circulation:+insights+into+the+neointimal+algorithm+and+management+strategies.">Mechanisms of vein graft adaptation to the arterial circulation: insights into the neointimal algorithm and management strategies.</a> Circulation Journal : Official Journal of the Japanese Circulation Society. 74, 1501-1512 (2010).</li><li>Tao, 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=A+simplified+murine+intimal+hyperplasia+model+founded+on+a+focal+carotid+stenosis.">A simplified murine intimal hyperplasia model founded on a focal carotid stenosis.</a> The American Journal of Pathology. 182, 277-287 (2013).</li><li>Berard, X., 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=Salvage+treatment+for+venous+aneurysm+complicating+vascular+access+arteriovenous+fistula:+use+of+an+exoprosthesis+to+reinforce+the+vein+after+aneurysmorrhaphy.">Salvage treatment for venous aneurysm complicating vascular access arteriovenous fistula: use of an exoprosthesis to reinforce the vein after aneurysmorrhaphy.</a> European Journal of Vascular and Endovascular Surgery : the Official Journal of the European Society for Vascular Surgery. 40, 100-106 (2010).</li></ol>

作者什么都没有透露。

这项研究揭示了体外静脉灌注系统（每股内含价值）在人类静脉进行大量的血流动力学研究。这个系统允许根据在没有通过体内循环细胞所释放的加重炎症和生长因子定义的血流动力学参数隐静脉灌注。因此，它提供了更好的理解涉及IH在人静脉移植物5,11,12,15的控制的基本途径。 可重复和可定量的血液动力学扰动体内限定。几个复杂的鼠显...

心血管疾病是发病率和死亡率在西方国家1的主要原因。尽管血管内治疗取得了进步，搭桥手术仍然是现代治疗的主体，从而五十多万元静脉移植在美国每年进行的。然而，尽管几十年的研究，下肢静脉移植30-60％范围内的最初几年，由于内膜增生（1H）2失败。机械力，特别是低切应力（SS）和高壁张力，关键是在这个增生反应3,4的启动和发展。为了解决这个问题，一个离体血管灌注系统（EVPS）中产生的学习，严格控制的血流动力学条件（压力和剪切应力），人隐静脉的行为下。在这项研究中，下列插入到动脉样循环，高压（平均= 100毫米汞柱）足以刺激prolif关合作和平滑肌细胞进入血管内膜层（1H）迁移。 哺乳动物的研究表明使用外部加固作为一种有效的方法来支持"静脉动脉化"，并抵消急性血液动力学改变静脉面临一次植入动脉周围环境。网状防止过 ​​度膨胀，增大的剪切应力，并降低了壁张力，因而IH 6-10。然而，潜在的机制和其适用于人类静脉改善旁路通畅性没有得到充分的表征。我们EVPS用于比较，在条件模拟的静脉面临一次插入动脉方案的改变（高剪切应力和压力），在外部大孔聚酯管状网的不存在和存在的大隐静脉的行为。防止病理性重构和IH，网格提供了其潜在的临床有效率11证据</sup&gt;。 这项研究1）引入体外控制下的压力和剪切应力2）人类隐静脉灌注模型显示，外部大孔涤纶网眼降低IH和提供了其潜在的临床应用至关重要的信息。

<table border="0" cellpadding="0" cellspacing="0" width="1113">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:453px;">Name</td>
			<td style="width:371px;">Company</td>
			<td style="width:145px;">Catalog Number</td>
			<td style="width:144px;">Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">RPMI 1640 - Glutamax</td>
			<td>Life Technologies</td>
			<td>61870-010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;"><a name="RANGE!A3">Penicilline/Streptomycine/Fungizone</a></td>
			<td>Bioconcept</td>
			<td>4-02F00-H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dextran from Leuconostoc spp. 500 gr.</td>
			<td>Sigma-Aldrich</td>
			<td>31390</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tampon PBS CHUV pH 7.1-7.3 1 lt.</td>
			<td>Laboratorium und Grosse Apotheke Dr. G. Bichsel AG</td>
			<td>100 0 324 00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cryosectionning embedding medium - Tissue-Tek OCT Compound</td>
			<td>Fisher Scientific</td>
			<td>14-373-65</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Silicon Tubing (Peroxide) L/S 16 (96400-16 ) - 7.5m</td>
			<td>Idex Health &#38; Science GMBH</td>
			<td>MF0037ST</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Y-splitter&#160;</td>
			<td>Idex Health &#38; Science GMBH</td>
			<td>Y-connector</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">35 mm Culture dish</td>
			<td>Sigma-Aldrich</td>
			<td>CLS430165-100EA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15 ml Falcon tube</td>
			<td>BD Bioscence</td>
			<td>352096</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">50 ml Falcon tube</td>
			<td>BD Bioscence</td>
			<td>352098</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Gearing pump - Reglo-Z</td>
			<td>Idex Health &#38; Science GMBH</td>
			<td>SM 895&#160;</td>
			<td>&#160;App-Nr 03736-00194</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pump Head</td>
			<td>Idex Health &#38; Science GMBH</td>
			<td>MI0008&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Monitoring Kit TRANSPAC IV</td>
			<td>icumedical</td>
			<td>011-0J736-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">20 mL Syringes</td>
			<td>B. Braun Medical SA</td>
			<td>4612041-02</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Etibon 3-0 FS-2</td>
			<td>Ethicon- Johnson&#38;Johnson</td>
			<td>EH7346H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mesh ProVena 6-8mm</td>
			<td>B. Braun Medical SA</td>
			<td>1105012-14</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NaCl: Sodium Chlorure Solution perfusion 0.9% (100 ml)</td>
			<td>B. Braun Medical SA</td>
			<td>534534</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Masterflex L/S Standard Drive</td>
			<td>Cole-Parmer Instrument Co</td>
			<td>7521-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Acquisition card</td>
			<td>National Instruments</td>
			<td>PCI-6024 E</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Flowmeter module</td>
			<td>Transonic Systems Inc.</td>
			<td>TS410 and T402</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stopcock with 3-ways</td>
			<td>BD Connexta Luerlock</td>
			<td>394600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Millex Filter</td>
			<td>Milian</td>
			<td>SE2M229I04</td>
			<td></td>
		</tr>
	</tbody>
</table>

procedure for human saphenous veins ex vivo perfusion and external reinforcement

现代疗法的广泛闭塞性动脉疾病的主要方法是静脉旁路移植术。然而，它的耐久性受到威胁内膜增生（1H），最终导致血管闭塞和移植失败。的机械力，特别是在低剪切应力和高壁张力，被认为是发起和维持这些细胞和分子变化，但其确切的贡献仍有待解开。以选择性地评估的压力和剪切应力IH的生物学上的作用， 离体灌注系统（EVPS）已创建灌注下动脉方案的大隐静脉（高剪切应力和高压力）的片段。进一步的技术创新，使两部分的同时灌注同出一脉，一个是外部的网状钢筋。脉中使用非接触式技术收获并立即在EVPS转移到实验室进行组装。新鲜ISOL的一个片段ATED静脉未灌注（对照组，第0天）。另外两个节段灌注长达7天，一个是完全为4毫米（直径）外部网的庇护。压力，流速，和脉冲速率连续地监测并调节以模仿血液动力学条件盛行的股动脉。在灌注完成后，静脉被拆卸，并用于组织学和分子分析。下离体条件下，高压灌注（动脉，平均= 100毫米汞柱）足以产生IH和人类静脉的重塑。这些改变都减小外部聚酯丝网的存在。

的每股内含价值提供了一个有价值的工具，以独立评估血流动力学的力量对人类隐静脉移植重建和IH。 图1示出了灌注腔和静脉支撑。在图1A和B，前静脉载体（ 图1A）和之后（ 图1B）组件，分别被描绘。它是由（从顶部至底部）的1平纹不锈钢管尺寸为9厘米，这可作为2为L形件，可以容易地滑动（从左至右）的载?...

Watch this Scientific Journal Video about Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement at JoVE.com

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

导致内膜增生（IH）和静脉移植物衰竭的发展机制仍然知之甚少。这项研究描述了一种体外系统灌注在控制流量和压力的人脉。另外外部钢筋网的效率限制IH的发展进行了评价。

步骤人类隐静脉<em&gt;前体内</em&gt;灌注和外部强化

The mainstay of contemporary therapies for extensive occlusive arterial disease is venous bypass graft. However, its durability is threatened by intimal ...

procedure-for-human-saphenous-veins-ex-vivo-perfusion-external

Research

JoVE Journal

Medicine

Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

11.1K 次观看.  Brigham and Women's Hospital/Harvard Medical School. 导致内膜增生（IH）和静脉移植物衰竭的发展机制仍然知之甚少。这项研究描述了一种体外系统灌注在控制流量和压力的人脉。另外外部钢筋网的效率限制IH的发展进行了评价。 

视频: Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

The success of liver transplantation has resulted in a dramatic organ shortage. In most transplant regions 20-30% of patients on the waiting list for liver transplantation die without receiving an organ transplant or are delisted for disease progression. One strategy to increase the donor pool is the utilization of marginal grafts, such as fatty livers, grafts from older donors, or donation after cardiac death (DCD). The current preservation technique of cold static storage is only poorly tolerated by marginal livers resulting in significant organ damage. In addition, cold static organ storage does not allow graft assessment or repair prior to transplantation.
These shortcomings of cold static preservation have triggered an interest in warm perfused organ preservation to reduce cold ischemic injury, assess liver grafts during preservation, and explore the opportunity to repair marginal livers prior to transplantation. The optimal pressure and flow conditions, perfusion temperature, composition of the perfusion solution and the need for an oxygen carrier has been controversial in the past.
In spite of promising results in several animal studies, the complexity and the costs have prevented a broader clinical application so far. Recently, with enhanced technology and a better understanding of liver physiology during ex vivo perfusion the outcome of warm liver perfusion has improved and consistently good results can be achieved.
This paper will provide information about liver retrieval, storage techniques, and isolated liver perfusion in pigs. We will illustrate a) the requirements to ensure sufficient oxygen supply to the organ, b) technical considerations about the perfusion machine and the perfusion solution, and c) biochemical aspects of isolated organs.

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts

Marginal grafts, such as fatty livers, grafts from older donors, or livers retrieved after cardiac death (DCD) tolerate conventional, cold static storage only poorly. We developed a novel model of subnormothermic ex vivo liver perfusion for preservation, assessment, and repair of marginal liver grafts prior to transplantation.

Subnormothermic的技术<em&gt;前体内</em&gt;肝脏灌注的存储，评估和修复边际移植肝

The success of liver transplantation has resulted in a dramatic organ shortage. In most transplant regions 20-30% of patients on the waiting list for ...

科研

医学

An Ex vivo Model to Study Hormone Action in the Human Breast

The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial cells tend to lose hormone receptor expression. Widely used hormone receptor positive breast cancer cell lines are of limited relevance to the in vivo situation. Here, we describe an ex vivo model to study hormone action in the human breast. Fresh human breast tissue specimens from surgical discard material such as reduction mammoplasties or mammectomies are mechanically and enzymatically digested to obtain tissue fragments containing ducts and lobules and multiple stromal cell types. These tissue microstructures kept in basal medium without growth factors preserve their intercellular contacts, the tissue architecture, and remain hormone responsive for several days. They are readily processed for RNA and protein extraction, histological analysis or stored in freezing medium. Fluorescence activated cell sorting (FACS) can be used to enrich for specific cell populations. This protocol provides a straightforward, standard approach for translational studies with highly complex, varied human specimens.

An Ex vivo Model to Study Hormone Action in the Human Breast

We have developed a novel ex vivo model to study hormone action in the human breast. It is based on tissue microstructures isolated from surgical breast tissue specimens which preserve tissue architecture, intercellular interactions, and paracrine signaling.

一个<em&gt;体外</em&gt;模型在乳腺癌研究激素的作用

The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial ...

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a serious problem. Therefore, the acceptance criteria for organ donors have been extended leading to the usage of marginal kidney grafts. These marginal organs tolerate cold storage poorly resulting in increased preservation injury and higher rates of delayed graft function. To overcome the limitations of cold storage, extensive research is focused on alternative normothermic preservation methods.
Ex vivo normothermic organ perfusion is an innovative preservation technique. The first experimental and clinical trials for ex vivo lung, liver, and kidney perfusions demonstrated favorable outcomes.
In addition to the reduction of cold ischemic injury, the method of normothermic kidney storage offers the opportunity for organ assessment and repair. This manuscript provides information about kidney retrieval, organ preservation techniques, and isolated ex vivo normothermic kidney perfusion (NEVKP) in a porcine model. Surgical techniques, set up for the perfusion solution and the circuit, potential assessment options, and representative results are demonstrated.

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to &#160;transplantation.

常温<em&gt;离体</em&gt;肾脏的灌注肾的移植保护移植前

Kidney transplantation has become a well-established treatment option for patients with end-stage renal failure. The persisting organ shortage remains a ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, normothermic ex vivo liver perfusion (NEVLP) has been introduced as a method to evaluate and modify organ function. NEVLP has many advantages in comparison to hypothermic and subnormothermic perfusion including reduced preservation injury, restoration of normal organ function under physiologic conditions, assessment of organ performance, and as a platform for organ repair, remodeling, and modification. Both murine and porcine NEVLP models have been described. We demonstrate a rat model of NEVLP and use this model to show one of its important applications &#8212; the use of a therapeutic molecule added to liver perfusate. Catalase is an endogenous reactive oxygen species (ROS) scavenger and has been demonstrated to decrease ischemia-reperfusion in the eye, brain, and lung. Pegylation has been shown to target catalase to the endothelium. Here, we added pegylated-catalase (PEG-CAT) to the base perfusate and demonstrated its ability to mitigate liver preservation injury. An advantage of our rodent NEVLP model is that it is inexpensive in comparison to larger animal models. A limitation of this study is that it does not currently include post-perfusion liver transplantation. Therefore, prediction of the function of the organ post-transplantation cannot be made with certainty. However, the rat liver transplant model is well established and certainly could be used in conjunction with this model. In conclusion, we have demonstrated an inexpensive, simple, easily replicable NEVLP model using rats. Applications of this model can include testing novel perfusates and perfusate additives, testing software designed for organ evaluation, and experiments designed to repair organs.

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury.

体外常温肝灌注的小动物模型

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Determining and Controlling External Power Output During Regular Handrim Wheelchair Propulsion

The use of a manual wheelchair is critical to 1% of the world&#39;s population. Human powered wheeled mobility research has considerably matured, which has led to improved research techniques becoming available over the last decades. To increase the understanding of wheeled mobility performance, monitoring, training, skill acquisition, and optimization of the wheelchair-user interface in rehabilitation, daily life, and sports, further standardization of measurement set-ups and analyses is required. A crucial stepping-stone is the accurate measurement and standardization of external power output (measured in Watts), which is pivotal for the interpretation and comparison of experiments aiming to improve rehabilitation practice, activities of daily living, and adaptive sports. The different methodologies and advantages of accurate power output determination during overground, treadmill, and ergometer-based testing are presented and discussed in detail. Overground propulsion provides the most externally valid mode for testing, but standardization can be troublesome. Treadmill propulsion is mechanically similar to overground propulsion, but turning and accelerating is not possible. An ergometer is the most constrained and standardization is relatively easy. The goal is to stimulate good practice and standardization to facilitate the further development of theory and its application among research facilities and applied clinical and sports sciences around the world.

Accurate and standardized assessment of external power output is crucial in the evaluation of physiological, biomechanical, and perceived stress, strain, and capacity in manual wheelchair propulsion. The current article presents various methods to determine and control power output during wheelchair propulsion studies in the laboratory and beyond.

在常规手部轮椅推进过程中确定和控制外部功率输出

The use of a manual wheelchair is critical to 1% of the world&#39;s population. Human powered wheeled mobility research has considerably matured, which ...

External Cephalic Version: Is it an Effective and Safe Procedure?

External cephalic version (ECV) is an effective procedure for reducing the number of cesarean sections. To date, there is no video publication showing the methodology of this procedure. The main objective is to show how to perform ECV with a specific protocol with tocolysis before the procedure and analgesia. Moreover, we describe and analyze the factors associated with successful ECV, and also compare to deliveries in the general pregnant population.
A retrospective and descriptive analysis of ECV carried out at the Hospital Clinico Universitario Virgen de la Arrixaca in Murcia (Spain) between 1/1/2014 and 12/31/2018 was assessed. The latest data available of labor deliveries in the local center, which is the biggest maternity department in Spain, were from 2018.
320 patients were recruited and 3 pregnant women were lost during the study. ECV was carried out at 37&#177;3 weeks gestation. ECV was successful in 82.5% (N=264). 19 complications were reported (5.9%): 8 vaginal bleeding (2.5%), 9 fetal bradycardia (2.8%), 1 preterm rupture of membranes (0.3%) and 1 cord prolapse (0.3%). A previous vaginal delivery increases the success rate of ECV ORadjusted=3.03 (1.62-5.68). Maternal Body Mass Index (BMI) affects the success of ECV ORadjusted=0.94 (0.89-0.99). Patients with BMI&#62;40 kg/m2 have an ORadjusted=0.09 (0.009-0.89) compared with those with BMI &#60;25 kg/m2. If ECV was successful, the cesarean delivery index is 22.2% (17.5-27.6%), the eutocic delivery index is 52.1% (46.1-58.1%) and the instrumented vaginal delivery index is 25.7% (20.7-31.2%). There are no differences in cesarean and eutocic delivery indexes after successful ECV. However, a successful ECV is associated with a 6.29% increase in the instrumented delivery rate (OR=1.63).
ECV is an effective procedure to reduce the number of cesarean sections for breech presentations. Maternal BMI and previous vaginal delivery are associated with ECV success. Successful ECV does not modify the usual delivery pattern.

This article shows how to perform the external cephalic version (ECV) by two experienced obstetricians in the obstetric operating room in the presence of an anesthesiologist and a midwife. The ECV is carried out with analgesia and tocolysis. Two attempts are made under ultrasonography control.

外部脑积水版本:它是一个有效和安全的程序吗？

External cephalic version (ECV) is an effective procedure for reducing the number of cesarean sections. To date, there is no video publication showing the ...

Normothermic Ex Vivo Pancreas Perfusion for the Preservation of Pancreas Allografts before Transplantation

Pancreas transplantation (PTx) is a curative treatment for people who live with the burden of a diagnosis of diabetes mellitus (DM). However, due to organ shortages and increasing numbers of patients being listed for PTx, new strategies are needed to increase the number of available grafts for transplantation. 
Static cold storage (SCS) is considered the gold standard for standard criteria organs. However, standard criteria donors (SCD) are becoming scarce and new strategies that can increase the rate of organ acceptance from extended criteria donors (ECD) are urgently needed. 
Normothermic ex vivo perfusion (NEVP) is one of the strategies that has become increasingly popular over the past couple of decades. This preservation method has already been used successfully in other organs (liver, kidneys, and lungs) but has been minimally explored in pancreas transplantation. The few papers that describe the method for pancreas show little success, edema being one of the major issues. The following manuscript describes the successful NEVP method and setup developed by our group to perfuse swine pancreas.

Normothermic Ex Vivo Pancreas Perfusion for the Preservation of Pancreas Allografts before Transplantation

Normothermic ex vivo machine perfusion (NEVP) has scarcely been explored for the preservation of pancreas allografts. We present an innovative preservation technique for pancreas allografts before transplantation.

非原生离体胰腺灌注用于移植前胰腺同种异 体 移植物的保存

Pancreas transplantation (PTx) is a curative treatment for people who live with the burden of a diagnosis of diabetes mellitus (DM). However, due to organ ...

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

One of the challenges in retina research is studying the cross-talk between different retinal cells such as retinal neurons, glial cells, and vascular cells. Isolating, culturing, and sustaining retinal neurons in vitro have technical and biological limitations. Culturing retinal explants may overcome these limitations and offer a unique ex vivo model to study the cross-talk between various retinal cells with well-controlled biochemical parameters and independent of the vascular system. Moreover, retinal explants are an effective screening tool for studying novel pharmacological interventions in various retinal vascular and neurodegenerative diseases such as diabetic retinopathy. Here, we describe a detailed protocol for retinal explants' isolation and culture for an extended period. The manuscript also presents some of the technical problems during this procedure that may affect the desired outcomes and reproducibility of the retinal explant culture. The immunostaining of the retinal vessels, glial cells, and neurons demonstrated intact retinal capillaries and neuroglial cells after 2 weeks from the beginning of the retinal explant culture. This establishes retinal explants as a reliable tool for studying changes in the retinal vasculature and neuroglial cells under conditions that mimic retinal diseases such as diabetic retinopathy.

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

This protocol presents and describes steps for the isolation, dissection, culturing, and staining of retinal explants obtained from an adult mouse. This method is beneficial as an ex vivo model for studying different retinal neurovascular diseases such as diabetic retinopathy.

成年小鼠视网膜的视网膜外植体作为研究视网膜神经血管疾病的离 体 模型

One of the challenges in retina research is studying the cross-talk between different retinal cells such as retinal neurons, glial cells, and vascular ...

<meta charset="utf8"></head><body>在大鼠 EVLP 模型中使用基于 PolyhHb 的灌注推进肺移植

Exploring Alternative Perfusion Solutions Using Next-Generation Polymerized Hemoglobin-Based Oxygen Carriers in a Model of Rat Ex Vivo Lung Perfusion

Lung transplantation is hampered by the lack of suitable donors. Previously, donors that were thought to be marginal or inadequate were discarded. However, new and exciting technology, such as ex vivo lung perfusion (EVLP), offers lung transplant providers extended assessment for marginal donor allografts. This dynamic assessment platform has led to an increase in lung transplantation and has allowed providers to use donors that were previously discarded, thus expanding the donor pool. Current perfusion techniques use cellular or acellular perfusates, and both have distinct advantages and disadvantages. Perfusion composition is critical to maintaining a homeostatic environment, providing adequate metabolic support, decreasing inflammation and cellular death, and ultimately improving organ function. Perfusion solutions must contain sufficient protein concentration to maintain appropriate oncotic pressure. However, current perfusion solutions often lead to fluid extravasation through the pulmonary endothelium, resulting in inadvertent pulmonary edema and damage. Thus, it is necessary to develop novel perfusion solutions that prevent excessive damage while maintaining proper cellular homeostasis. Here, we describe the application of a polymerized human hemoglobin (PolyhHb)-based oxygen carrier as a perfusate and the protocol in which this perfusion solution can be tested in a model of rat EVLP. The goal of this study is to provide the lung transplant community with key information in designing and developing novel perfusion solutions, as well as the proper protocols to test them in clinically relevant translational transplant models.

<meta charset="utf8"></head><body>作者聚焦：利用下一代聚合人血红蛋白改善大鼠供体肺的评估和保存

Here, we describe the application of a polymerized human hemoglobin (PolyhHb)-based oxygen carrier as a perfusate and the protocol in which this perfusion solution can be tested in a model of rat ex vivo lung perfusion.