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>

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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

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型:教训制定鼠EVLP课程中学

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

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 ...

Human Dupuytren's Ex Vivo Culture for the Study of Myofibroblasts and Extracellular Matrix Interactions

Organ fibrosis or &#8220;scarring&#8221; is known to account for a high death toll due to the extensive amount of disorders and organs affected (from cirrhosis to cardiovascular diseases). There is no effective treatment and the in vitro tools available do not mimic the in vivo situation rendering the progress of the out of control wound healing process still enigmatic.
To date, 2D and 3D cultures of fibroblasts derived from DD patients are the main experimental models available. Primary cell cultures have many limitations; the fibroblasts derived from DD are altered by the culture conditions, lack cellular context and interactions, which are crucial for the development of fibrosis and weakly represent the derived tissue. Real-time PCR analysis of fibroblasts derived from control and DD samples show that little difference is detectable. 3D cultures of fibroblasts include addition of extracellular matrix that alters the native conditions of these cells. 
As a way to characterize the fibrotic, proliferative properties of these resection specimens we have developed a 3D culture system, using intact human resections of the nodule part of the cord. The system is based on transwell plates with an attached nitrocellulose membrane that allows contact of the tissue with the medium but not with the plastic, thus, preventing the alteration of the tissue. No collagen gel or other extracellular matrix protein substrate is required. The tissue resection specimens maintain their viability and proliferative properties for 7 days. This is the first &#8220;organ&#8221; culture system that allows human resection specimens from DD patients to be grown ex vivo and functionally tested, recapitulating the in vivo situation.

Human Dupuytren's Ex Vivo Culture for the Study of Myofibroblasts and Extracellular Matrix Interactions

Dupuytren&#8217;s disease (DD) is a fibroproliferative disease of the palm of the hand. Here, we present a protocol to culture resection specimens from DD in a three-dimensional (3D) culture system. Such short-term culture system allows preservation of the 3D structure and molecular properties of the fibrotic tissue.

人掌腱膜挛缩的<em&gt;离体</em&gt;文化为肌成纤维细胞的研究和细胞外基质相互作用

Organ fibrosis or &#8220;scarring&#8221; is known to account for a high death toll due to the extensive amount of disorders and organs affected (from ...

Ex Situ Normothermic Machine Perfusion of Donor Livers

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous perfusion of organs provides the opportunity to improve organ quality and allows ex situ viability assessment of donor livers prior to transplantation. This video article provides a step by step protocol for ex situ normothermic machine perfusion (37 &#176;C) of human donor livers using a device that provides a pressure and temperature controlled pulsatile perfusion of the hepatic artery and continuous perfusion of the portal vein. The perfusion fluid is oxygenated by two hollow fiber membrane oxygenators and the temperature can be regulated between 10 &#176;C and 37 &#176;C. During perfusion, the metabolic activity of the liver as well as the degree of injury can be assessed by biochemical analysis of samples taken from the perfusion fluid. Machine perfusion is a very promising tool to increase the number of livers that are suitable for transplantation.

Ex Situ Normothermic Machine Perfusion of Donor Livers

Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.

供肝易地常温灌注机

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous ...

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 ...

Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing

The use of patient-derived xenografts for modeling cancers has provided important insight into cancer biology and drug responsiveness. However, they are time consuming, expensive, and labor intensive. To overcome these obstacles, many research groups have turned to spheroid cultures of cancer cells. While useful, tumor spheroids or aggregates do not replicate cell-matrix interactions as found in vivo. As such, three-dimensional (3D) culture approaches utilizing an extracellular matrix scaffold provide a more realistic model system for investigation. Starting from subcutaneous or intracranial xenografts, tumor tissue is dissociated into a single cell suspension akin to cancer stem cell neurospheres. These cells are then embedded into a human-derived extracellular matrix, 3D human biogel, to generate a large number of microtumors. Interestingly, microtumors can be cultured for about a month with high viability and can be used for drug response testing using standard cytotoxicity assays such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and live cell imaging using Calcein-AM. Moreover, they can be analyzed via immunohistochemistry or harvested for molecular profiling, such as array-based high-throughput kinomic profiling, which is detailed here as well. 3D microtumors, thus, represent a versatile high-throughput model system that can more closely replicate in vivo tumor biology than traditional approaches.

Patient-derived xenografts of glioblastoma multiforme can be miniaturized into living microtumors using 3D human biogel culture system. This in vivo-like 3D tumor assay is suitable for drug response testing and molecular profiling, including kinomic analysis.

Microtumors使用3D人体Biogel培养体系和病人源性胶质母细胞瘤的Kinomic剖析和药物反应测试的生成

The use of patient-derived xenografts for modeling cancers has provided important insight into cancer biology and drug responsiveness. However, they are ...

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, ...

Optocardiography and Electrophysiology Studies of Ex Vivo Langendorff-perfused Hearts

Small animal models are most commonly used in cardiovascular research due to the availability of genetically modified species and lower cost compared to larger animals. Yet, larger mammals are better suited for translational research questions related to normal cardiac physiology, pathophysiology, and preclinical testing of therapeutic agents. To overcome the technical barriers associated with employing a larger animal model in cardiac research, we describe an approach to measure physiological parameters in an isolated, Langendorff-perfused piglet heart. This approach combines two powerful experimental tools to evaluate the state of the heart: electrophysiology (EP) study and simultaneous optical mapping of transmembrane voltage and intracellular calcium using parameter sensitive dyes (RH237, Rhod2-AM). The described methodologies are well suited for translational studies investigating the cardiac conduction system, alterations in action potential morphology, calcium handling, excitation-contraction coupling and the incidence of cardiac alternans or arrhythmias.

The objective of this study was to establish a method for investigating cardiac dynamics using a translational animal model. The described experimental approach incorporates dual-emission optocardiography in conjunction with an electrophysiological study to assess electrical activity in an isolated, intact porcine heart model.

前维沃·兰根多夫注入心脏的心电生理学研究

Small animal models are most commonly used in cardiovascular research due to the availability of genetically modified species and lower cost compared to ...

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 ...

步骤人类隐静脉

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Subnormothermic的技术

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供肝易地常温灌注机

常温

Microtumors使用3D人体Biogel培养体系和病人源性胶质母细胞瘤的Kinomic剖析和药物反应测试的生成

体外常温肝灌注的小动物模型

前维沃·兰根多夫注入心脏的心电生理学研究

在常规手部轮椅推进过程中确定和控制外部功率输出