这项工作是由1R03DK096151的部分资助。

所有小鼠购自杰克逊实验室（巴港，ME）购买并根据美国国立卫生研究院的准则和科罗拉多州丹佛IACUC大学的批准饲养的无病原体的条件下，在美国科罗拉多州丹佛市，芭芭拉·戴维斯中心动物设施的大学。 1.体供肾收获<ol><li>消毒所有仪器，整个过程中戴上无菌手套，并保持无菌区。完成所有的手术配合使用的手术显微镜。 </li><li>从手术戊巴比妥麻醉（60毫克/千克IP）捐助者拔下鼠标供肾。确定麻醉通过脚趾捏的深度，并观察呼吸频率。 </li><li>剪辑的皮毛，然后固定在鼠标的4路限制。准备用碘伏皮肤和悬垂鼠标以消毒方式。 </li><li>做2厘米的垂直中线腹部切口，进入腹腔。缩回肠优化和外部它到胸部。保持包装在无菌湿润纱布在整个过程中的肠。 </li><li>确定伟大的腹部血管和动员他们，找出任何腰椎分行，要么烧灼或用10/0尼龙线将它们连接起来。 </li><li>立即远端左肾血管挑逗下腔静脉（IVC）和腹主动脉（AA）除了钝性分离了2mm左右。结扎与10/0尼龙和分裂的小动脉和静脉分支的肾血管。 </li><li>现在分开肾动脉仔细肾静脉由这些结构钝性分离。这允许在肾静脉的近端的精确创建卡雷尔补丁，这将被用于形成一个端 - 侧吻合到收件人IVC。 </li><li>识别，结扎和划分左肾上腺血管。这允许访问肾上腺主动脉和6-0丝线缝合周围放置在主动脉中准备结扎，但不依赖在这个时候。 </LI&gt; <li>动员肾脏，血管和输尿管从周围的仪表板。旋转肾脏的权利和结扎/分任后支。然后肾脏返回到左侧。 </li><li>现在的注意力集中输尿管。而不会干扰肾门不受周围的仪表板照顾输尿管保留输尿管船只。划分输尿管在输精管的水平。肾脏是现在可以恢复。 </li><li>缓缓注入肝素进入远端下腔静脉，从而heparinizing捐赠的300台。阕6-0丝线缝合周围放置肾上腺AA，并通过远端腹主动脉与肝素盐水（100 U / ml）的0.8毫升灌注左肾。 </li><li>一旦灌注已停止创建静脉卡雷尔补丁立即消除回流到肾。缩回肾静脉朝向肾揭示了AA和肾动脉下方。 </li><li>分邻近肾动脉的主动脉创建脚跟，并如图1 C箍。从供体中取出肾脏，血管和输尿管。 </li><li>直接取出肾脏到预先制备的收件人（0分钟冷缺血时间）或储存在4℃下在所选择的一个预定时间中的溶液，直至植入的时间。安乐死的捐助者exsanguanation和颈椎脱位。恢复供肾的总时间大约为15-20分钟。 </li></ol> 2.肾脏植入技术<ol><li> （如果需要60毫克/公斤的IP初始剂量为25mg / kg的IP补充剂量）麻醉受体小鼠戊巴比妥。确定麻醉通过脚趾捏的深度，并观察呼吸频率。剪辑的皮毛，然后固定在鼠标的4路约束和应用眼药膏的眼睛。准备用碘伏皮肤和悬垂鼠标以消毒方式。 </li><li>做2厘米的垂直中线腹部切口，进入腹腔。缩回肠优和外部化到胸部。保持包装在无菌湿润纱布在整个过程中的肠。 </li><li>在这个时候进行正确的性肾切除术。结扎肾动脉和静脉用6-0丝线缝合。切除肾脏远端的缝线。结扎输尿管以6-0丝线，然后分近端肾。 </li><li>隔离腹主动脉和下腔静脉（IVC）以下的肾血管。周围放置主动脉4-0棉花的关系，下腔静脉优于然后逊于吻合部位。内场识别并结扎所有腰椎血管用10-0尼龙缝合。 </li><li>结棉关系，第一下后跟优越。在这种方式的一些血液被保持在使主动脉切开术更容易主动脉。 </li><li>形成具有30G针的主动脉切开术进入主动脉的内腔中。延长切口用细显微剪到2mm左右的长度。 </li><li>做一个结束捐助主动脉脚跟和脚趾袖侧吻合术收件人主动脉以下的方式。放置一个10-0尼龙缝合针留在供体主动脉和切口在收件人主动脉和领带的下角。 </li><li>将第二10-0尼龙相反的第一个供体主动脉和切口腹主动脉和领带的高级角落。做一个连续缝合线由优到劣主动脉的侧壁和领带对先前放置的逗留针。然后缝合内侧壁上的运行方式和领带。 </li><li>请停止供体肾静脉在以下时尚收件人下腔静脉端侧吻合。穿刺IVC用30G针和细微剪刀延长切口。领带供体肾静脉的切口下腔静脉的下角和10-0尼龙。使肾静脉和下腔静脉和领带之间的连续缝合线。 注：这也是非常重要的全层通过缝合针，包括血管adventit的ia和内膜得以实现。边缘外翻也确保有内膜到内膜接触，这有助于密封和所述吻合的愈合。虽然止血凝血剂可以减少渗漏有用，我们建议外科医生，而不是依靠良好的技术。 </li><li>确保吻合是“干净的”。也就是说，将针迹时的相对的壁没有被捕获。这将导致一个显著收缩流动，这将导致一个失败接枝和在极端情况下，以后肢麻痹。 注：另外一个非常重要的因素是确保吻合缝线的张力也处于最佳状态。过松又会有不可逆转的泄漏，过紧而狭窄的流动将导致。如果在动脉侧，这将导致在接枝的灌注差，如果在静脉侧的拥塞肾脏将导致。 </li><li>松开远端4-0棉领带重新建立静脉血流。一旦静脉止血美国吻合术已经观察到逐步放松的近端4-0棉结，观察动脉吻合术止血。 </li><li>从鼠标中取出棉关系，一旦两者吻合术被认为是安全的。皮尔斯用20G的针头创建两个孔膀胱。 </li><li>通过孔通过弯曲钳提示，并通过与输尿管的近端钳子膀胱拉施主输尿管被锚固到膀胱壁具有两个10-0尼龙缝合线。修剪输尿管从第二孔使输尿管向膀胱内缩回，并关闭两个10-0尼龙缝合线的孔突出的多余长度。 </li><li>返回肠腹部。关闭腹壁两层用5-0丝线缝合的运行方式。 </li><li>辖1.0毫升丸无菌，温暖的生理盐水到腹部为交易结束后的液体复苏，并注入0.8的生理盐水皮下手术后毫升。没有其他supporti在手术期间需要五个措施。 </li><li>恢复在加温毯动物。总植入时间约。 35-45分钟。施用镇痛剂，如丁丙诺啡，0.05毫克/公斤，SC 0.1-0.2毫升在过程的开始和每6-12小时72小时后的操作。 </li></ol> 3.对侧肾切除术<ol><li>根据协议要求，几天植入手术后，进行异氟烷麻醉下对侧肾切除术（5％异氟烷吸入诱导，维护1.5-2.0％）。 </li><li>进入腹腔，轻轻缩回肠到动物的右。揭露左肾和钝器解剖与周围筋膜。 </li><li>结扎肾动脉，静脉和输尿管用6-0丝线缝合，然后切除上面的缝线肾脏，取出肾脏。总手术时间约。 10分钟。 </li></ol> 4.移植评估<ol><li>测定血清肌酐9使用碱性苦味酸法（谢菲反应）。 </li></ol>

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The authors have nothing to disclose.

掌握这种移植技术是困难的，但一旦完成了它是一个非常强有​​力的研究工具。病人的外科医生/研究人员将通过注重细节和技术，这是在小动物模型的关键是掌握任何外科手术，更使一致性得到回报。掌握鼠标肾移植的技术难度大很多倍，而且极有可能在其他小动物移植模型中的经验，必须处理这个过程之前获得。 即，将针以保持一个专利管腔时的相对的血管壁的...

自1973年以来在小鼠肾脏移植模型一直是一个有价值的研究工具，但技术问题阻碍了它的广泛使用。多年来，一些论文已发表的细节改进/改进这一程序。作为主要血管实体器官移植模型这个过程可能是仅次于异位心脏移植模型，这也是在1973年6设计由罗素实验室。这两种型号借给自己的研究分为异体排斥反应，对移植肾功能延迟发展和局部缺血再灌注损伤。 其中最常见的问题与肾移植报是动脉血栓形成4,5,7，我们也经历了我们实验室的发病率相对较高。因此，我们开始着手进行血栓形成的文献综述，并可能发现这个技术问题的原因，也设计一个可能的解决方案。血栓形成的最可能的原因是血液需要从收件人主动脉，成体供肾肾主动脉，然后就到了供肾动脉有点曲折的道路。此路径将导致湍流，肾动脉，可导致血小板激活和血栓形成。根据近期的观察和搜索相关文献8-14中 ，我们提出了一个新的技术，减少血栓形成，以0％。 此处所描述的技术从在动脉踵和胎趾袖口哪个设施改善血液流的形成以前报道的技术而不同，显著减少血栓形成。袖带是通过将横跨肾动脉口的面的红外线肾主动脉在低于45°到主动脉（图1A＆1B）的纵向轴线的角度形成的。这导致了袖带大约2mm的长度。静脉卡雷尔补丁是由横断第r形成烯醛静脉进入下腔静脉，从而增加了封套的直径。该红外供肾腹主动脉脚跟和脚趾袖口端 - 端吻合于受体腹主动脉和供体肾静脉/下腔静脉修补程序端 - 端吻合给收件人腹部下腔静脉（IVC） 。输尿管随后被引入并如由韩等人3锚固在膀胱。 在这项研究中未经处理的移植，只有热缺血时间（ 即 ，不冷缺血）进行比较。在这种情况下，热缺血是指从血液流过供体肾脏的停止的时间（下面的步骤1.11），并在受者（下面步骤2.11）的接枝的再灌注。冷缺血是指肾脏未灌注和保存在冷藏直到植入程序开始的时间。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Instrument</td>
			<td>Roboz #</td>
			<td>Fine Science Tools #</td>
			<td>Arosurgical #</td>
		</tr>
		<tr>
			<td>Straight micro-dissecting forcep #5</td>
			<td>RS-5015</td>
			<td>11295-51</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved micro-dissecting forcep #7</td>
			<td>RS-5047</td>
			<td>11297-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved serrated forcep</td>
			<td>RS-5137</td>
			<td>11052-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Vannas micro-dissecting scissors, short</td>
			<td>RS-5610</td>
			<td></td>
			<td>09.140.08</td>
		</tr>
		<tr>
			<td colspan="2">Micro-dissecting scissors, straight, sharp, long</td>
			<td></td>
			<td>11.602.11</td>
		</tr>
		<tr>
			<td>Micro spring handle needle holder</td>
			<td></td>
			<td></td>
			<td>11.549.15</td>
		</tr>
		<tr>
			<td>Straight mosquito forcep</td>
			<td></td>
			<td>91308-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-dissecting scissors, straight, blunt</td>
			<td>RS-5962</td>
			<td>14078-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro-dissecting scissors, curved, blunt</td>
			<td>RS-5981</td>
			<td>14079-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro retractor</td>
			<td>RS-6540</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Instrument tray, 10&#8221; x 6 &#189;&#8221; x &#190;&#8221;</td>
			<td>RT-1350S</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Silk suture, 5/0, 22.5m spool</td>
			<td></td>
			<td>18020-50</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Suture</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10/0 nylon</td>
			<td></td>
			<td></td>
			<td>T4A10Q07</td>
		</tr>
		<tr>
			<td>5/0 silk</td>
			<td></td>
			<td></td>
			<td>E19A05N</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gloves</td>
			<td></td>
			<td>Drapes</td>
			<td></td>
		</tr>
		<tr>
			<td>Biogel from Medex Supply</td>
			<td></td>
			<td>Precept, #64-9012-9</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Syringes</td>
			<td></td>
			<td>Cotton applicators</td>
			<td></td>
		</tr>
		<tr>
			<td>B-D 1cc insulin, #329424</td>
			<td></td>
			<td colspan="2">Fisher-brand, #23-400-100</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone-Iodine swabs</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PDI, #B40600</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>4/0 Cotton ties</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td colspan="2">Domestic cotton autoclaved with instruments</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

murine kidney transplant technique

The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique using the donor aorta to form the arterial anastomosis instead of the renal artery was developed and reported in 1993 by Kalina and Mottram 2 with a further advancement coming from the same laboratory in 1999 3. While one can become proficient in this model, a search of the literature reveals that many labs still experience a high proportion of graft loss due to arterial thrombosis. We describe here a technique that was devised in our laboratory that vastly reduces the arterial thrombus reported by others 4,5. This is achieved by forming a heel-and-toe cuff of the donor infra-renal aorta that facilitates a larger anastomosis and straighter blood flow into the kidney.

这种手术方法使得无论是简单的移植物的存活/排斥反应的研究，还是蛮复杂的实验方案。在图中下面，我们展示了使用该改进的动脉吻合术的优点。使用这项技术，我们已经显著降低动脉血栓形成的发病率从35％至0％从而提高了生产率。我们已经用这种技术一年以上在保持相同的0％血栓形成的结果。 图1描述的方法中的动脉踵和胎趾袖口这是这种新技术的基础上的形成。这个袖口提?...

Watch this Scientific Journal Video about Murine Kidney Transplant Technique at JoVE.com

Murine Kidney Transplant Technique

The goal of this manuscript is to describe the steps required to perform a kidney transplant in a mouse, paying particular attention to the details of the arterial anastomosis.

小鼠肾脏移植技术

The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. Although it took some years for other labs to become ...

Research

JoVE Journal

Medicine

15.7K 次观看.  University of Colorado, Denver. The goal of this manuscript is to describe the steps required to perform a kidney transplant in a mouse, paying particular attention to the details of the arterial anastomosis. 

视频: Murine Kidney Transplant Technique

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize their blood glucose levels, islet transplantation has been proposed to be a potential treatment for type 1 diabetes 1,2. More recently, advances in human islet transplantation have further strengthened this view 1,3. However, two major limitations prevent islet transplantation from being a widespread clinical reality: (a) the requirement for large numbers of islets per patient, which severely reduces the number of potential recipients, and (b) the need for heavy immunosuppression, which significantly affects the pediatric population of patients due to their vulnerability to long-term immunosuppression. Strategies that can overcome these limitations have the potential to enhance the therapeutic utility of islet transplantation.
Islet transplantation under the mouse kidney capsule is a widely accepted model to investigate various strategies to improve islet transplantation. This experiment requires the isolation of high quality islets and implantation of islets to the diabetic recipients. Both procedures require surgical steps that can be better demonstrated by video than by text. Here, we document the detailed steps for these procedures by both video and written protocol. We also briefly discuss different transplantation models: syngeneic, allogeneic, syngeneic autoimmune, and allogeneic autoimmune.

Transplantation of isolated islets has been proposed to be a potential treatment for type 1 diabetes. Here we describe a method to isolate islets from mouse pancreata and transplant them to the subcapsular space of the kidney.

小鼠胰岛的分离和包膜下肾移植的方法

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

科研

医学

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

用改进的袖套法小鼠颈部心脏移植模型

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

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 ...

Murine Heterotopic Heart Transplant Technique

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique was developed and reported in 2001 by Niimi. Described here are the techniques that have evolved over more than a decade in the hands of three surgeons (Plenter, Grazia, Pietra) in our center. These techniques are now being passed on to a younger generation of surgeons and researchers.
Based largely on the Niimi experience, the procedures used have evolved in the fine details - details which we will endeavor to relate here in such a way that others may be able to use this very useful model. Like Niimi, we have found that a video aid to learning is a priceless resource for the beginner.

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

小鼠异位心脏移植技术

It is now over forty years since this technique was first reported by Corry, Wynn and Russell. Although it took some years for other labs to become ...

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. 
Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. 
Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

使用Active波尔图腔静脉分流术猪肝脏采购和原位移植技术

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation ...

Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis

Vascular rejection that leads to transplant arteriosclerosis (TA) is the leading representation of chronic heart transplant failure. In TA, the immune system of the recipient causes damage of the arterial wall and dysfunction of endothelial cells and smooth muscle cells. This triggers a pathological repair response that is characterized by intimal thickening and luminal occlusion. Understanding the mechanisms by which the immune system causes vasculature rejection and TA may inform the development of novel ways to manage graft failure. Here, we describe a mouse aortic interposition model that can be used to study the pathogenic mechanisms of vascular rejection and TA. The model involves grafting of an aortic segment from a donor animal into an allogeneic recipient. Rejection of the artery segment involves alloimmune reactions and results in arterial changes that resemble vascular rejection. The basic technical approach we describe can be used with different mouse strains and targeted interventions to answer specific questions related to vascular rejection and TA.

We describe a protocol for aortic interposition grafting in mice. The goal of the protocol is to provide a model with which to study pathological processes and therapeutic strategies relevant to alloimmune reactions in arteries and the resultant arterial changes that contribute to organ transplant failure.

的同种免疫引起的血管性排斥反应和移植物动脉硬化小鼠模型

Vascular rejection that leads to transplant arteriosclerosis (TA) is the leading representation of chronic heart transplant failure. In TA, the immune ...

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 ...

An &#8220;All-laser&#8221; Endothelial Transplant

The &#8220;all laser&#8221; assisted endothelial keratoplasty is a procedure that is performed with a femtosecond laser used to cut the donor tissue at an intended depth, and a near infrared diode laser to weld the corneal tissue. The proposed technique enables to reach the three main goals in endothelial keratoplasty: a precise control in the thickness of the donor tissue; its easy insertion in the recipient bed and a reduced risk of donor lenticule dislocation. The donor cornea thickness is measured in the surgery room with optical coherence tomography (OCT), in order to correctly design the donor tissue dimensions. A femtosecond laser is used to cut the donor cornea. The recipient eye is prepared by manual stripping of the descemetic membrane. The donor endothelium is inserted into a Busin-injector, the peripheral inner side is stained with a proper chromophore (a water solution of Indocyanine Green) and then it is pulled in the anterior chamber. The transplanted tissue is placed in the final and correct location and then diode laser welding is induced from outside the eyeball. The procedure has been performed on more than 15 patients evidencing an improvement in surgery performances, with a good recovery of visual acuity and a reduced donor lenticule dislocation event.

An &amp;#8220;All-laser&amp;#8221; Endothelial Transplant

An all laser procedure is proposed in the endothelial transplant. The surgical technique is based on the use of a femtosecond laser to prepare the donor tissue. Laser welding technique is then used to secure the donor endothelium in the correct position.

“全激光”内皮移植

The &#8220;all laser&#8221; assisted endothelial keratoplasty is a procedure that is performed with a femtosecond laser used to cut the donor tissue at an ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the availability of the widest range of molecular probes and reagents to perform in vivo as well as in vitro studies. Based on our experience with various murine transplantation models, we developed a heterotopic pancreas transplantation model in mice with the intent to analyze mechanisms underlying severe ischemia reperfusion injury-associated early graft damage. In contrast to previously described techniques using suture techniques, herein we describe a new procedure using a non-suture cuff technique. 
In recent years, we have performed more than 300 pancreas transplantations in mice with an overall success rate of &gt;90%, a success rate never described before in mouse pancreas transplantation. The backbone of this non-suture cuff technique for graft revascularization consists of two major steps: (I) pulling the recipient vessel over a polyethylene/polyamide cuff and fixing it with a circumferential ligature, and (II) placing the donor vessel over the everted recipient vessel and fixing it with a second circumferential ligature. The resultant continuity of the endothelial layer results in less thrombogenic lesions with high patency rates and, finally, high success rates.
In this model, arterial anastomosis is achieved by pulling the abdominal aorta of the donor graft over the everted common carotid artery of the recipient animal. Venous drainage of the graft is achieved by pulling the portal vein of the graft over the everted external jugular vein of the recipient. This manuscript provides details and crucial steps of the organ recovery and organ implantation procedures, which will allow researchers with microsurgical skills to perform the transplantation successfully in their laboratories.

Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages.

应用改良袖套技术的小鼠胰腺移植模型

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.
Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

使用改性非缝合铐技术的宫颈主动脉移植模型

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...