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><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>Micro-dissecting scissors, straight, sharp, long</td><td></td><td>11.602.11</td><td></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&amp;rdquo; x 6 &amp;frac12;&amp;rdquo; x &amp;frac34;&amp;rdquo;</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>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>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>Syringes</td><td></td><td>Cotton applicators</td><td></td></tr><tr><td>B-D 1cc insulin, #329424</td><td></td><td>Fisher-brand, #23-400-100</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>4/0 Cotton ties</td><td></td><td></td><td></td></tr><tr><td>Domestic cotton autoclaved with instruments</td><td></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?...

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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

16.0K 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 Renal Transplantation Procedure

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell et al over 30 years ago (1) and refined by Zhang et al years later (2), few people in the world have mastered this procedure. In our laboratory we have successfully performed 1200 orthotopic kidney transplantations with &gt; 90% survival rate. The key points for success include stringent control of reperfusion injury, bleeding and thrombosis, both during the procedure and post-transplantation, and use of 10-0 instead of 11-0 suture for anastomoses.
Post-operative care and treatment of the recipient is extremely important to transplant success and evaluation. All renal graft recipients receive antibiotics in the form of an injection of penicillin immediately post-transplant and sulfatrim in the drinking water continually. Overall animal health is evaluated daily and whole blood creatinine analyses are performed routinely with a portable I-STAT machine to assess graft function.

Renal transplantation in mice is a technically challenging procedure that requires careful post-operative care and treatment for success.

小鼠肾移植程序

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell ...

生物学

Isolation and Transplantation of Different Aged Murine Thymic Grafts.

The mechanisms that regulate the efficacy of thymic selection remain ill-defined. The method presented here allows in vivo analyses of the development and selection of T cells specific for self and foreign antigens. The approach entails implantation of thymic grafts derived from various aged mice into immunodeficient scid recipients. Over a relatively short period of time the recipients are fully reconstituted with T cells derived from the implanted thymus graft. Only thymocytes seeding the thymus at the time of isolation undergo selection and develop into mature T cells. As such, changes in the nature and specificity of the engrafted T cells as a function of age-dependent thymic events can be assessed. Although technical expertise is required for successful thymic transplantation, this method provides a unique strategy to study in vivo a wide range of pathologies that are due to or a result of aberrant thymic function and/or homeostasis.

This report provides a detailed description of transplanting murine thymi from different aged donor mice under the kidney capsule of immunodeficient mouse recipients. The goal of this approach is to model T cell development and thymic selection events in vivo.

隔离和不同的老年小鼠胸腺移植的移植。

The mechanisms that regulate the efficacy of thymic selection remain ill-defined. The method presented here allows in vivo analyses of the development and ...

免疫与感染

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

鼠肾移植:移植排斥反应模型

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Fate Mapping of Human Embryonic Stem Cells by Teratoma Formation 

Human embryonic stem cells (hESCs) have an unlimited capacity for self-renewal, and the ability to differentiate into cells derived from all three embryonic germ layers (1). Directed differentiation of hESCs into specific cell types has generated much interest in the field of regenerative medicine (e.g., (2-5)), and methods for determining the in vivo fate of selected or manipulated hESCs are essential to this endeavor. We have adapted a highly efficient teratoma formation assay for this purpose. A small number of specifically selected hESCs is mixed with undifferentiated wild type hESCs and Phaseolus vulgaris lectin to form a cell pellet. This is grafted beneath the kidney capsule in an immunodeficient mouse. As few as 2.5 x 105 hESCs are needed to form a 16 cm3 teratoma within 8-12 weeks. The fate of the originally selected hESCs can then be determined by immunohistochemistry. This method provides a valuable tool for characterizing tissue-specific reagents for cell-based therapy.

Fate Mapping of Human Embryonic Stem Cells by Teratoma Formation

Directed differentiation of hESCs into specific cells has generated much interest in regenerative medicine. We provide a concise, step-by-step protocol for determining the in vivo fate of selected hESCs that provides a valuable tool for characterizing tissue-specific reagents for cell-based therapy.

人类胚胎干细胞畸胎瘤形成命运的映射

Human embryonic stem cells (hESCs) have an unlimited capacity for self-renewal, and the ability to differentiate into cells derived from all three ...

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality of life when compared to dialysis. Although the last few decades have seen major improvements in patient outcomes following kidney transplantation, the increasing shortage of available organs represents a severe problem worldwide. To expand the donor pool, marginal kidney grafts recovered from extended criteria donors (ECD) or donated after circulatory death (DCD) are now accepted for transplantation. To further improve the postoperative outcome of these marginal grafts, research must focus on new therapeutic approaches such as alternative preservation techniques, immunomodulation, gene transfer, and stem cell administration.
Experimental studies in animal models are the final step before newly developed techniques can be translated into clinical practice. Porcine kidney transplantation is an excellent model of human transplantation and allows investigation of novel approaches. The major advantage of the porcine model is its anatomical and physiological similarity to the human body, which facilitates the rapid translation of new findings to clinical trials. This article offers a surgical step-by-step protocol for an autotransplantation model and highlights key factors to ensure experimental success. Adequate pre- and postoperative housing, attentive anesthesia, and consistent surgical techniques result in favorable postoperative outcomes. Resection of the contralateral native kidney provides the opportunity to assess post-transplant graft function. The placement of venous and urinary catheters and the use of metabolic cages allow further detailed evaluation. For long-term follow-up studies and investigation of alternative graft preservation techniques, autotransplantation models are superior to allotransplantation models, as they avoid the confounding bias posed by rejection and immunosuppressive medication.

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

异位自体肾移植的猪模型:有步骤，分步协议

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

Production of Humanized Mouse via Thymic Renal Capsule Grafting, CD34+ Cells Injection, and Cytokine Delivery

Animal models provide a vital translation between in vitro and in vivo biomedical research. Humanized mouse models provide a bridge in the representation of human systems, thereby allowing for a more accurate study of pathogenesis, biomarkers, and many other scientific queries. In this method described, immune-deficient NOD-scid IL2R&#947;null (NSG) mice are implanted with autologous thymus, injected with liver-derived CD34+ cells followed by a series of injected cytokine deliveries. In contrast to other models of a similar nature, the model described here promotes an improved reconstitution of immune cells by delivering cytokines and growth factors via transgenes encoded in AAV8 or pMV101 DNA-based vectors. Moreover, it offers long-term stability with reconstituted mice having an average lifespan of 30 weeks after CD34+ injections. Through this model, we hope to provide a stable and impactful method of studying immunotherapy and human disease in a murine model, thus demonstrating the need for predictive preclinical models.

<meta charset="utf8"></head><body>通过胸腺肾包膜移植、CD34 + 细胞注射和细胞因子递送生产人源化小鼠

Humanized mouse models provide a more accurate representation of the human immune microenvironment. This manuscript describes the process in which these models are created through a renal graft of human thymus, injection of human CD34+ cells, and the targeted delivery of human cytokine transgenes to promote CD34+ cell proliferation and differentiation.

通过胸腺肾包膜移植、CD34 + 细胞注射和细胞因子递送生产人源化小鼠

Animal models provide a vital translation between in vitro and in vivo biomedical research. Humanized mouse models provide a bridge in the representation ...

A Modified Surgical Technique for Kidney Transplantation in Mice

Kidney transplantation in mice is a complicated and challenging surgery procedure. There are very few publications demonstrating the key steps of this operation. Therefore, this article introduces the technique and points out the surgical caveats associated with this operation. In addition, important modifications in comparison to the conventional procedure are demonstrated. Firstly, a patch of the abdominal aorta is cut and prepared so that the proximal bifurcations of the renal artery, including the ureteral artery are transected together with the donor kidney en bloc. This reduces the risk of a ureter necrosis and avoids the development of a urinary tract occlusion. Secondly, a new method of the vascular anastomosis is demonstrated that allows the operator to flexibly increase or decrease the size of the anastomosis after renal transplant reperfusion has already been initiated. This avoids the development of vessel strictures and intraabdominal bleeding. Thirdly, a technique that enables the anastomosis of the delicate donor ureter and the recipient bladder that does not cause a trauma is shown. Adopting this protocol can shorten the operation time and reduces the damage to the recipient's bladder, thereby significantly increasing the operation success rate for the recipient mice.

This protocol presents a new surgical technique of mouse kidney transplantation focusing on a modified arterial anastomosis strategy. A vascular suture technique including a simple and safer ureter-bladder anastomosis method is also presented. These modifications shorten the operation time and improve the success rate of the mouse kidney transplantation procedure.

一种用于小鼠肾移植的改良手术技术

Kidney transplantation in mice is a complicated and challenging surgery procedure. There are very few publications demonstrating the key steps of this ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of renal replacement therapy. Over the past few decades, the rat kidney transplantation model has been used to study the immunological phenomena of rejection and tolerance. This model has become an indispensable tool to test new immunomodulatory pharmaceuticals and regimens prior to proceeding with expensive preclinical large animal studies.
This protocol provides a detailed overview of how to reliably perform orthotopic kidney transplantation in rats. This protocol includes three distinctive steps that increase the probability of success: perfusion of the donor kidney by flushing through the portal vein and the use of a cuff system to anastomose the renal veins and ureters, thereby decreasing cold and warm ischemia times. Using this technique, we have achieved survival rates beyond 6 months with normal serum creatinine in animals with syngeneic or tolerant kidney transplants. Depending on the aim of the study, this model can be modified by pre- or posttransplant treatments to study the acute, chronic, cellular, or antibody-mediated rejection. It is a reproducible, reliable, and cost-effective animal model to study different aspects of kidney transplantation.

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

大鼠原位肾移植: 一种新的简化的手术方法

Kidney transplantation offers increased survival rates and improved quality of life for patients with end-stage renal disease, as compared to any type of ...