这项工作是由澳大利亚资助计划- 1029205国家健康和医学研究委员会（NHMRC）的支持，由澳大利亚Microsearch基金会，由Myee科德林顿医学研究基金会。

在此之前开始实验，获得规划批准的实验从相关机构的动物护理伦理委员会。在按照你的机构的要求，保持小鼠。下面的协议已获得悉尼大学，皇家阿尔弗雷德王子医院委员会。 在此之前开始手术，所有仪器应彻底清洗，并浸泡在80％酒精消毒。一些机构规定蒸压然而，这可以在长期内损害精细的显微外科器械。尽可能使用一次性无菌设备。 1，麻醉<ol><li>麻醉小鼠异氟醚在密闭容器内，然后将其放置在仰卧的工作板，快速连接麻醉鼻锥。麻醉捏后脚，以确保没有反射试验是否足够。 </li><li>剃的皮肤用手术刀片和消毒用80％的乙醇。另外，在70％异丙醇2％氯己定，可用于皮肤杀菌。在麻醉诱导的异氟醚浓度为3％，但减少到1-1.5％进行维护。作出轻微调整，这个浓度，以维持正常呼吸和心脏速率。维持动物的加热板上的温度。 </li></ol> 2，供体手术<ol><li>使开胸通过切割胸部通过胸廓两侧从肋缘至腋下随后在剑突的水平横向切割，以形成胸壁皮瓣，露出心脏和血管。提起皮瓣头来旁边钉选到操作板。撕下心包，暴露心脏和血管。 </li><li>提起下腔静脉钳，一手通过下腔静脉其它注射1ml冷肝素生理盐水近端到心脏，然后把一小动脉夹上腔静脉，以防止措施的灌注液反向通过针孔吨流动。 </li><li>用纱布和棉签，收回心脏向下，显露升主动脉和肺动脉。通过一对microscissors中的一个叶片通过通道（横窦）后到主动脉和肺动脉束并切断主动脉和肺动脉一起尽可能远侧尽可能地确保足够的长度用于吻合术。 </li><li>领带和划分下腔静脉，右侧上腔静脉（SVC），用6-0丝线离开SVC和肺静脉。另配合下腔静脉和右SVC然后将一个线程后到心脏与肺静脉环绕左SVC在一起，配合它。通过切割血管远端的关系，然后将其在冷的无菌盐水中储存于4℃直至移植的供体部位收获心脏。这会导致供体放血死亡。 </li></ol> 3，受体手术<ol><li>麻醉收件人如上面（第1节）。仔细剃腹部，以避免刺激，然后做一个通过剖腹手术从耻骨正中切口，以剑突和收回使用回形针弯曲形成拉钩。包裹肠在温暖的无菌生理盐水浸泡的纱布和缩回到腹部的右上方。 </li><li>揭露基础设施肾主动脉和下腔静脉，主动脉自由和下腔静脉从左侧肾动脉和静脉捆绑段至髂分叉使用烧灼设备从他们的腰椎血管分裂。注意使烧灼是在适当的温度和所用足够的时间来划分和密封该容器端部。 </li><li>应用无创小血管夹到主动脉和下腔静脉段远端和近端。由第一穿刺用30号针头的孔做一个切口在主动脉的前壁;然后切一个切口垂直microscissors相匹配的供体主动脉的大小。冲洗主动脉管腔与肝素生理盐水，以消除任何BLOOD血栓。 </li><li>在供体心脏带给受区覆盖着纱布浸泡在冷盐水，并将其放置到右侧腹部。确保供体主动脉定位在接受​​者主动脉与供体肺动脉位于旁边的收件人IVC旁切口。 </li><li>供体主动脉端 - 侧吻合首先使用运行10-0尼龙缝线从近侧转角和缝合沿左侧开始的收件人主动脉直到到达远端的角落，在该阶段的动物是通过180°的旋转。轻轻的供体心脏移动到腹部的左侧以暴露右侧的主动脉，并通过右侧从远端到近端主动脉壁的继续缝合。收盘前主动脉吻合，轻轻冲洗管腔与肝素生理盐水，以消除任何血块和空气。 </li><li>吻合供体肺动脉（PA）的端至端与接收方腔静脉。做一个切口版tically与下腔静脉在一个站点根据动脉吻合前壁microscissors。吻合的捐赠者PA到收件人IVC与运行从下腔静脉管腔内左侧壁的远端开始10-0尼龙缝线。在到达近端后，继续沿前右侧壁到前端的缝线来完成吻合术。在关闭吻合术，轻轻冲洗管腔以除去任何凝块和空气。 </li><li>释放前用棉签血管钳，地方明胶海绵块周围的吻合部位，并轻轻向下按压直到止血的实现。在再血管化的时候，先松开远侧钳，随后近侧钳。 </li><li>血运重建后，在37℃适用温生理盐水的移植外，协助其恢复。接枝通常会立即开始颤动，并自发地恢复为窦性心律在几分钟之内。注入0.6毫升温暖皮下注射生理盐水，以维持收件人的水合作用。手术前完成注入丁丙诺啡皮下镇痛</li><li>关闭腹部伤口用一个5-0可吸收缝线运行两个图层。首先，完成内层和沿皮肤继续。 </li></ol> 4，恢复和移植物监测<ol><li>注射氨苄青霉素感染的预防和放置在一个加热垫收件人在37℃进行恢复。大多数动物迅速恢复，通常喝，往往在3个小时吃。如果鼠标显示窘迫征象，仔细检查，以确定病因。如果没有明显的原因，治疗与丁丙诺啡和密切监测。请教兽医，如果症状严重或持续时间超过8小时，并考虑安乐死。如果有必要，继续给予丁丙诺啡12小时注射，直到症状消失。如果鼠标显示困难的症状8小时后，给予持续12丁丙诺啡每小时注射，直到他们解决。咨询兽医，如果症状持续时间超过48小时，如果需要安乐死。 </li><li>监测移植心脏的跳动直接腹部触诊，记录节拍的强度+ + + +为​​一个健康的移植到+为弱拍，由于先进的排斥反应及 - 由于完成的排斥移植的不跳动。每日监测小鼠在第10天，然后3次，每周为实验的持续时间。 </li></ol>

<ol>
	<li>Aramaki, O., 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=Interleukin-10+but+not+transforming+growth+factor-beta+is+essential+for+generation+and+suppressor+function+of+regulatory+cells+induced+by+intratracheal+delivery+of+alloantigen.">Interleukin-10 but not transforming growth factor-beta is essential for generation and suppressor function of regulatory cells induced by intratracheal delivery of alloantigen.</a> Transplantation. 79, 568-576 (2005).</li><li>Chen, R. H., Bushell, A., Fuggle, S. V., Wood, K. J., Morris, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+granzyme+A+and+perforin+in+mouse+heart+transplants+immunosuppressed+with+donor-specific+transfusion+and+anti-CD4+monoclonal+antibody.">Expression of granzyme A and perforin in mouse heart transplants immunosuppressed with donor-specific transfusion and anti-CD4 monoclonal antibody.</a> Transplantation. 61, 625-629 (1996).</li><li>Poulin, L. 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=Interleukin-9+promotes+eosinophilic+rejection+of+mouse+heart+allografts.">Interleukin-9 promotes eosinophilic rejection of mouse heart allografts.</a> Transplantation. 76, 572-577 (2003).</li><li>Ono, K., Lindsey, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+technique+of+heart+transplantation+in+rats.">Improved technique of heart transplantation in rats.</a> J. Thorac. Cardiovasc. Surg. 57, 225-229 (1969).</li><li>Corry, R. J., Winn, H. J., Russell, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Primarily+vascularized+allografts+of+heart+in+mice.">Primarily vascularized allografts of heart in mice.</a> Transplantation. 16, 343-350 (1973).</li><li>Larsen, C. 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=CD40-gp39+interactions+play+a+critical+role+during+allograft+rejection.+Suppression+of+allograft+rejection+by+blockade+of+the+CD40-gp39+pathway.">CD40-gp39 interactions play a critical role during allograft rejection. Suppression of allograft rejection by blockade of the CD40-gp39 pathway.</a> Transplantation. 61, 4-9 (1996).</li><li>Saitovitch, D., Bushell, A., Mabbs, D. W., Morris, P. J., Wood, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+induction+of+transplantation+tolerance+with+a+nondepleting+anti-Cd4+monoclonal+antibody+and+donor-specific+transfusion+before+transplantation.+A+critical+period+of+time+is+required+for+development+of+immunological+unresponsiveness.">Kinetics of induction of transplantation tolerance with a nondepleting anti-Cd4 monoclonal antibody and donor-specific transfusion before transplantation. A critical period of time is required for development of immunological unresponsiveness.</a> Transplantation. 61, 1642-1647 (1996).</li><li>Callaghan, C. J., 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=Regulation+of+allograft+survival+by+inhibitory+FcgammaRIIb+signaling.">Regulation of allograft survival by inhibitory FcgammaRIIb signaling.</a> J. Immunol. 189, 5694-5702 (2012).</li><li>Qian, 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=Impact+of+donor+MHC+Class+I+or+Class+II+antigen+deficiency+on+first-+and+second-set+rejection+of+mouse+heart+or+liver+allografts.">Impact of donor MHC Class I or Class II antigen deficiency on first- and second-set rejection of mouse heart or liver allografts.</a> Immunology. 88, 124-129 (1996).</li><li>Wang, 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=Spontaneous+acceptance+of+mouse+kidney+allografts+is+associated+with+increased+Foxp3+expression+and+differences+in+the+B+and+T+cell+compartments.">Spontaneous acceptance of mouse kidney allografts is associated with increased Foxp3 expression and differences in the B and T cell compartments.</a> Transpl. Immunol. 24, 149-156 (2011).</li><li>Liu, F., Kang, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+Heart+Transplantation+in+Mice.">Heterotopic Heart Transplantation in Mice.</a> J. Vis. Exp. , (2007).</li><li>Mottram, P. L., 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=Electrocardiographic+monitoring+of+cardiac+transplants+in+mice.">Electrocardiographic monitoring of cardiac transplants in mice.</a> Cardiovasc. Res. 22, 315-321 (1988).</li></ol>

作者什么都没有透露。

小鼠心脏移植是需要相当大的手术技巧掌握一个具有挑战性的显微外科方法。最具挑战性的方面是小直径的血管。此外，有必要限制接收者操作时间和出血。该技术对小鼠心脏移植首先由Corry 等人所述。在1973年，后经莫特拉姆等人 12。我们的修改包括以下几点。首先，供体心脏冷肝素盐水下列开胸主动脉和肺动脉束灌注后不久和横断立即灌注防止热缺血期间收获。其次，从?...

器官移植的动物模型可以提供有价值的信息，以提高治疗的临床移植的患者。小鼠模型是研究器官移植排斥或接受免疫机制，由于大范围的转基因小鼠和具体的小鼠不适用于其他动物模型1-3试剂特别有用。与移植小鼠模型的一个挑战是捐助者和受助人的需要相当的技术技能，以达到满意的结果的小尺寸。 一种技术是在大鼠4其后由科里等 5适用于小鼠心脏移植首先描述了心中的异位移植。这项技术涉及的准备之前，供体手术接受者和无灌注供体心脏，这两者都可能损害心脏移植或t的生存他收件人。该过程已被广泛地用作最初描述检查移植排斥和耐受6-8的机制。其他人已经适应了原有的大鼠心脏移植Ono和林赛的心脏移植在小鼠9,10过程。最近，一种技术被发布给小鼠心脏移植是克服了与Corry 等人 11的方法相关联的问题。在这里描述的协议采用了修改，基于莫特拉姆等人 12的方法，该方法包括: 在原位灌注冷肝素生理盐水开胸后立即和事先执行操作供体到受体手术，以减少接受者操作时间。此外，我们使用非创伤性小血管夹，而不是6-0真丝领带。虽然血管夹有采取更多的空间的缺点，他们更容易比的关系来控制，这不应该是太紧或太松并且是那么简单，除去比夹。我们的方法是使用一个单一的连续缝合为容器吻合，虽然在最初的学习阶段的替代方法是使用缝线留在近侧和远侧角，以确保缝合线的均匀性，因此该吻合的通畅。

<table><tbody><tr><td>Operating microscope&amp;nbsp;</td><td>Leica, Heerbrugg, Switzerland</td><td>M651</td><td>10-25X magnification</td></tr><tr><td>Anesthetic machine&amp;nbsp;</td><td>Vet Quip Pty Ltd, Sydney, Australia</td><td>Vett3</td><td>Capable of delivering a mixture of isoflurane and oxygen in air&amp;nbsp;</td></tr><tr><td>Operating board&amp;nbsp;</td><td>Hardware store or office supplier</td><td></td><td>Dense cork or synthetic capable of taking pins</td></tr><tr><td>Heparinized saline (5 U/ml, 4 &amp;deg;C)</td><td>Pfizer, USA</td><td>FW25</td><td>In 1 ml syringe with 23 G needle on ice</td></tr><tr><td>Normal saline (37 &amp;deg;C)</td><td>AstraZeneca Pty Ltd, Australia</td><td>4538</td><td>In 1 ml syringe with 23 G needle on warm pad</td></tr><tr><td>Sutures 10- nylon, 5-0 Vicryl</td><td>Ethicon, Inc. NJ, USA</td><td>2870G/J421H</td><td></td></tr><tr><td>Buprenorphine (0.05 mg/kg in 0.1 ml saline)</td><td>Reckitt Benckiser, Sydney, Australia</td><td></td><td>In 1 ml syringe with 23 G needle on ice</td></tr><tr><td>Ampcillin (5 mg/kg in 0.1 ml saline)</td><td>Aspen, Sydney, Australia</td><td></td><td>In 1 ml syringe with 23 G needle on ice</td></tr><tr><td>Gel Foam</td><td>Pharmacia &amp;amp; Upjohn Co. USA</td><td>801289304</td><td>Cut into small pieces</td></tr><tr><td>High temperature cautery device</td><td>Medtronic, USA</td><td>8444000</td><td></td></tr><tr><td>Heating Pad/Right Temp</td><td>Kent Scientific Corporation, Turrington, CT 06790</td><td></td><td></td></tr><tr><td>&lt;strong&gt;Microsurgery instruments:&lt;/strong&gt;</td><td>Shanghai Medical Instruments Co. Ltd.,&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Microneedleholders</td><td>Shanghai, China</td><td>WT2020</td><td></td></tr><tr><td>Microscissors</td><td>&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;</td><td>WT1020</td><td></td></tr><tr><td>Microforceps (straight tip)</td><td>&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;</td><td>WA3010</td><td></td></tr><tr><td>Microforceps (curved tip)</td><td>&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;</td><td>WA3020</td><td></td></tr><tr><td>Micromosquito clamps (1 pair)</td><td>&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;</td><td>W40350</td><td></td></tr><tr><td>Microvessel atraumatic clamps (1 pair)</td><td>&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp; &quot;</td><td>W40130/W40150</td><td></td></tr></tbody></table>

a modified method for heterotopic mouse heart transplantion

老鼠经常被用来作为心脏移植供，受者在移植免疫学研究，由于大范围的转基因小鼠和试剂可用的。的困难，提出由于动物的体积小，参与心脏移植的显微外科的相当大的技术挑战。特别是，移植术后早期高企的技术故障可能导致收件人死亡和术后并发症，如后肢瘫痪或无跳动的心脏。在这里，完整的技术异位小鼠心脏移植证实，涉及收获捐赠的心脏而其随后植入到受体小鼠。供体心脏收获立即就地灌注在与升主动脉和肺动脉的冷肝素生理盐水和横断。接受者操作涉及制备腹主动脉和下腔静脉（IVC），其次是端 - 端吻合术使用一个单一的运行10-0 microsuture和捐助肺动脉与受体下腔静脉的吻合类似的捐助主动脉与受体主动脉。以下操作的动物被注射用0.6 ml生理盐水皮下注射和允许恢复在37℃下加热垫。从227小鼠心脏移植手术的结果总结，成功率在48小时的86.8％。在48小时的13.2％的失败，5（2.2％）出现后肢麻痹，10（4.4％）有因移植缺血性损伤和/或血栓形成非跳动的心脏，15（6.6％），48小时内死亡。

经过最初的培训期，227例小鼠异位心脏移植在我们的小组进行了分析。在最初24小时内，成功率为90.3％，并且在48小时是86.8％。 48小时内的30（13.2％）的故障，5（2.2％）出现后肢瘫痪，不得不被施以安乐死，10（4.4％）有非跳动的心脏，由于移植物缺血损伤和/或血栓形成，而15 （6.6％），48小时内死亡。一些实验接枝残余示于表1的各种应变的组合。同一供体和受体菌株之间的心脏移?...

Watch this Scientific Journal Video about A Modified Method for Heterotopic Mouse Heart Transplantion at JoVE.com

A Modified Method for Heterotopic Mouse Heart Transplantion

A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

对于异位小鼠心脏移植技术的一种改进方法

Mice are often used as heart transplant donors and recipients in studies of transplant immunology due to the wide range of transgenic mice and reagents ...

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12.0K 次观看.  University of Sydney. A technique is demonstrated for the microsurgical procedure for heterotopic transplantation of hearts in mice, including simplified methods for donor harvesting and recipient vessel anastomosis.

视频: A Modified Method for Heterotopic Mouse Heart Transplantion

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable for studying rejection and immune response now that newer transgenic and gene knockout mice are available, and a large number of immunologic reagents have been developed. The heart transplant model is less stringent than the skin transplant models, although technically more challenging. We have developed a modified technique and have completed over 1000 successful cases of heterotopic heart transplantation in mice. When making anastomosis of the ascending aorta and abdominal aorta, two stay sutures are placed at the proximal and distal apexes of recipient abdominal aorta with the donor s ascending aorta, then using 11-0 suture for anastomosis on both side of aorta with continuing sutures. The stay sutures make the anastomosis easier and 11-0 is an ideal suture size to avoid bleeding and thrombosis.
When making anastomosis of pulmonary artery and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s pulmonary artery. The left wall of the inferior vena cava and donor s pulmonary artery is closed with continuing sutures in the inside of the inferior vena cava after, one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s pulmonary artery are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

小鼠异位心脏移植

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

科研

生物学

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 ...

医学

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

颈椎异位心脏移植小鼠

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

一种新的显微模型异位，整块胸壁，胸腺，和心脏移植小鼠

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

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

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

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

大鼠异位心脏和心肌细胞移植的免疫模型

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

免疫与感染

Optimization of the Cuff Technique for Murine Heart Transplantation

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to improve surgical efficiency. In our lab, we have optimized the cuff technique with two major steps. First, we used the inner tube technique to insert a temporary inner tube into the external jugular vein and carotid artery blood vessel to facilitate eversion of the vessel over the cuff. Second, we performed complete heterotopic cardiac transplantation through the collaboration of two experienced surgeons. These modifications effectively reduced the operation time to 25 minutes, with a success rate of 95%. In this report, we describe these procedures in detail and provide a supplemental video. We believe that this report on the improved cuff technique will offer practical guidance for murine heterotopic heart transplantation and will enhance the utility of this mouse model for basic research.

We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time.

穆林心脏移植袖口技术的优化

Murine cardiac transplantation has been performed for more than 40 years. With advancements in microsurgery, certain new techniques have been used to ...

Mouse Heterotopic Cervical Cardiac Transplantation Utilizing Vascular Cuffs

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after transplantation, and the impact of immunomodulatory therapies on graft rejection. Heterotopic cervical heart transplantation in mice was first described in 1991 using sutured anastomoses and subsequently modified to include cuff techniques. This modification allowed for improved success rates, and since then, there have been multiple reports that have proposed further technical improvements. However, translation into more widespread utilization remains limited due to the technical difficulty associated with graft anastomoses, which requires precision to achieve adequate length and caliber of the cuffs to avoid vascular anastomotic twisting or excessive tension, which can result in damage to the graft. The present protocol describes a modified technique for performing heterotopic cervical cardiac transplantation in mice which involves cuff placement on the recipient's common carotid artery and the donor's pulmonary artery in alignment with the direction of the blood flow.

Mouse cardiac transplantation models represent valuable research tools for studying transplantation immunology. The present protocol details mouse heterotopic cervical cardiac transplantation that involves the placement of cuffs on the recipient's common carotid artery and the donor's pulmonary artery trunk to allow for laminar blood flow.

利用血管袖带的小鼠异位宫颈心脏移植

Murine models of cardiac transplantation are frequently utilized to study ischemia-reperfusion injury, innate and adaptive immune responses after ...

A Modified Murine Heterotopic Heart Transplant Protocol Matching Contemporary Standards of Aseptic Technique, Anesthesia, and Analgesia

The development of experimental models of cardiac transplantation in animals has contributed to many advances in the fields of immunology and solid organ transplantation. While the heterotopic vascularized murine cardiac transplantation model was initially utilized in studies of graft rejection using combinations of mismatched inbred mouse strains, access to genetically modified strains and therapeutic modalities can provide powerful new preclinical insights. Fundamentally, the surgical methodology for this technique has not changed since its development, especially with respect to important factors such as aseptic technique, anesthesia, and analgesia, which make material impacts on postsurgical morbidity and mortality. Additionally, improvements in perioperative management are expected to provide improvements in both animal welfare and experimental outcomes. This paper reports upon a protocol developed in collaboration with a subject matter expert in veterinary anesthesia and describes the surgical technique with an emphasis on perioperative management. Additionally, we discuss the implications of these refinements and provide details on troubleshooting critical surgical steps for this procedure.

The present paper describes a modified technique for heterotopic vascularized cardiac transplantation with updated aseptic technique, analgesia, and anesthesia.

符合当代无菌技术、麻醉和镇痛标准的改良小鼠异位心脏移植方案

The development of experimental models of cardiac transplantation in animals has contributed to many advances in the fields of immunology and solid organ ...

Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the established technique for a modified blood circuit reconstruction in a heterotopic abdominal heart transplantation model is presented. This method uses the intrathoracic inferior vena cava (IIVC) instead of the pulmonary artery of the donor heart for the anastomosis to the inferior vena cava of the recipient. It is facilitating and improving success rates for abdominal heart transplantation in mice.

A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated.

腹部小鼠心脏移植模型中的血液回路重建

The surgical technique of heterotopic abdominal heart transplantation in mice is a standard model for research in transplantation immunology. Here, the ...

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for investigating the mechanism of cardiac allograft rejection in preclinical studies because of their high homology with human genes. This understanding would help develop unique approaches to improving patients&#39; long-term survival treated with cardiac allografts. In a mouse model, abdominal donor heart implantation is commonly performed with an end-to-end&#160;anastomosis to the recipient&#39;s aorta and inferior vena cava using stitches. In this model, the donor&#39;s heart is implanted by end-to-end anastomosis to the recipient&#39;s carotid artery and jugular vein by the modified-Cuff technique. The transplantation surgery is performed without stitching and thus may increase the survival of the recipient since there is no interference with the blood supply and venous reflux of the lower body. This mouse model would help investigate the mechanisms underlying the immunological and pathological (acute/chronic) rejection of cardiac allografts.

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

一种改进的袖带技术用于小鼠颈椎异位心脏移植模型

Cardiac allograft rejection limits the long-term survival of patients after heart transplantation. A mouse heart transplantation model is ideal for ...