这项工作是由健康研究和心脏及中风BC与育空地区的基金会（JCC）的加拿大学院资助。

本研究所有协议进行了审查并批准了西蒙·弗雷泽大学的动物保健伦理委员会。使用的Balb / CYJ（H2 D）供体小鼠和C57BL / 6（H2 B）受体小鼠检查异体反应。小鼠被用于8到12周之间的年龄的实验。请使用雌性或雄性小鼠。同基因控制包括从C57BL / 6捐助者主动脉段到C57BL / 6收件人。 1，供体和受体准备注意:无论是捐赠者和接受者麻醉和术前准备，以尽量减少移植缺血。注射麻醉剂中使用的协议，以防止阻塞的动物通过所需吸入麻醉剂的递​​送设备。然而，如果需要的话，吸入麻醉剂是合适的替代方案。从最初的注射的麻醉药，整个过程需要大约90分钟来完成。移植物的缺血时间小于THAñ30分钟。 <ol><li>麻醉小鼠用氯胺酮的腹膜内注射（100mg / kg的10毫克/毫升）和甲苯噻嗪（10毫克/千克; 1毫克/毫升）。鼠标将在10至15分钟服用镇静剂。按捏动物垫脚的脂肪部分评估麻醉深度。施用麻醉剂鸡尾酒的原剂量的1/3，根据需要，直到动物未表现出撤回反射。监测呼吸频率每鸡尾酒给予密切了。手术过程中，通过用钳子捏的前腹壁评估麻醉水平，每15分钟。将小鼠保持深度麻醉，60到90分钟。 </li><li>润滑鼠标的眼睛眼药膏，以防止干燥，而在麻醉下。 </li><li>剃头发尽可能靠近皮肤​​尽可能对腹部腹侧区域从中间胸部到耻骨。要特别小心不要尼克任何肤质。不要使用脱毛霜，因为它可能被吸收我扎成的皮肤和可能是炎症。 </li><li>发生在仰卧位动物在加热表或垫上面一条毛巾。 </li><li>与2％氯己初步刷洗干净的手术部位。准备一个大的工作区，以最大限度地提高手术视野。开始洗涤在手术部位的中心，并移动到外侧的线性或环状方式。处置纱布。重复此过程至少为5次以上。应用酒精消毒片到同一地区。一旦酒精是干的，准备与优碘溶液和悬垂性用无菌纱布清洁区。 </li></ol> 2.终端到终端的过程吻合<ol><li>供体手术<ol><li>保持整个操作无菌技术。清洁使用前的所有台面和手术台上表面与0.5％加速过氧化氢溶液。包装和高温灭菌所有手术器械，纱布，在使用前的窗帘和礼服。验证仪器的使用无菌蒸汽灭菌指示条放置在每个包。无菌手术手套的使用和处置的手术之间。对于多次手术，消毒使用的手术器械与热玻璃珠灭菌。 </li><li>将供体小鼠在一个干净的，薄的有机玻璃板包裹着手术显微镜的放大倍数8-30X在无菌的悬垂性仰卧位。 </li><li>在确保在1.1节所列足够的手术麻醉，进行手术治疗。 </li><li>用无菌剪刀，使一个单一的中线下腹部纵切口，从耻骨到xyphoid过程。 </li><li>用一个小拉钩，打开腹腔壁暴露腔。 </li><li>用消毒棉签，轻轻收回肠优越于动物的左，盖上纱布蘸生理盐水。移动生殖器官下方并找到下腹主动脉和下腔静脉（IVC）。 MOIS10暴露的组织定期用生理盐水溶液。 </li><li>使用医疗5号镊子，从下腔静脉分离主动脉，从左侧肾动脉分叉的水平。用10-0尼龙单丝缝合线结扎小树枝附近的主动脉。 </li><li>一旦捐赠者的主动脉已经脱离了下腔静脉，饱和容器用生理盐水，覆盖外露的腔体与湿纱布留出供体到无菌区。检查供体（呼吸道和心血管功能，以及麻醉深度），每15分钟的状态。开始对接受者操作，如下所述（步骤2.2.1至2.2.7）分离主动脉。 </li><li>一旦收件人主动脉已经脱离了下腔静脉和预留，返回显微镜下的捐助。交叉夹紧（近端和远端到感兴趣的段）的供体主动脉，大约5毫米的间隔，用两个4毫米微血管夹具。 </li><li>使用Vannas-蒂宾根microscissors，横切腹主动脉的一个小的移植物区段（3至4毫米的长度）。 </li><li>使用地下25 5/8针连接到注射器，冲洗切除主动脉肝素（100U / ml）的生理盐水溶液。确保针的尖端不会接触到与容器接触。 </li><li>在肝素地方主动脉上冰（100 U / ml）的盐溶液，备用。虽然仍处于深度麻醉，松开夹子微血管。安乐死供体放血。 </li><li>在切除的30分钟植入供体容器。虽然有可能通过切除主动脉的较大长度，以用于多个收件人一个供体容器中，保持缺血时间为小于30分钟。 </li></ol></li><li>收件人操作<ol><li>保持无菌技术在整个操作中，如供体手术。 </li><li>将受体小鼠在薄薄的有机玻璃板包裹着手术显微镜的放大倍数8-30X在无菌的悬垂性仰卧位。 </li><li>自动对焦之三确保足够的麻醉，进行手术时，动物不会出现撤退反射。 </li><li>用无菌剪刀，使一个单一的中线下腹部纵切口，从耻骨到xyphoid过程。 </li><li>用一个小拉钩，打开腹腔壁暴露腔。 </li><li>用消毒棉签，轻轻收回肠优越于动物的左，盖上纱布蘸生理盐水。移动生殖器官下方并找到下腹主动脉和下腔静脉。用生理盐水滋润定期暴露组织。 </li><li>从下腔静脉从左侧肾动脉分叉的水平分离主动脉。如有必要，可使用聚酰胺10-0缝合丝结扎接近主动脉小树枝。 </li><li>交叉夹紧（近端和远端到感兴趣的段）的主动脉，大约5毫米的间隔，用两个4毫米微血管夹具。 </li><li>使用Vannas-杜宾microscissors，使单个水平主动脉切开术和切除腹主动脉的一小部分（不大于0.5mm），以适应供体主动脉移植物。 </li><li>冲洗切除主动脉肝素（100U / ml）的生理盐水溶液。供体主动脉移植物应该是适当的长度连接收件人的主动脉横断两端。 </li><li>放置在原位位置供体主动脉移植物和吻合捐赠者的移植物年底到收件人的结束，匹配相应的解剖移植方向与收件人。 </li><li>轻轻抓住容器中膜外耳炎和外翻稍微使用医疗五号钳子。使用镊子，驱动通过血管壁的整个厚度附着到10-0聚酰胺单纤维缝合线的针，以确保供体主动脉移植到收件人的切除容器中。请注意，以确保船舶开口没有封闭到期牛逼容器的后壁邻无意缝线。 </li><li>对于连续的线圈，放置停留缝线在上部和下部的移植物的端部9点钟。起始于从3点钟的接枝的上端，吻合的切除端部与2运行缝合并固定缝线中止缝合。 <ol><li>翻转移植，并继续运行缝合背血管的一部分，符合原产地停留缝合。确保不施加太多的压力在容器上。重复该缝合用于下吻合。 注意:间断缝合开始相同的方式连续缝合，不同的是将容器吻合与停留缝线之间的三个分开的线圈。吻合时间通常为20分钟。 </li></ol></li><li>一旦吻合术完成后，松开远端微血管钳允许逆行血液流和检查的部位吻合泄漏。如果有泄漏，立即出现Ÿ放置一针一针关闭缺陷部位。如果没有出血的部位，然后释放近端夹。 </li><li>检查移植，检查有没有血梗阻的移植，以及近端和收件人的血管远端部分。在这两个供体和收件人的血管剧烈脉冲模式是一个主要迹象表明，血液畅通无阻。使用钳子，轻轻抓住逗留缝线的一端，并略微外翻容器来检查容器的后壁。 注:应该没有褶皱血管壁的的部位吻合两端。除去夹子后血流不畅是血栓形成的标志。 </li><li>用棉签，返回肠道进入腹腔。 </li><li>随着柄持针钳和镊子格雷夫，关闭腹壁采用连续缝合5-0聚丙烯缝线。用相同的关闭皮层采用皮内缝合关闭。 </li><li>移植完成后立即施用Torbugesic（1毫克/千克）即时。 </li><li>马上给的顺序，阿替美唑（1毫克/千克），酮洛芬（5毫克/千克），温热乳酸林格氏液皮下注射。 </li><li>马上手术后，放置在老鼠在水毯子过夜（12小时）加热笼。在麻醉恢复期间，单独放置动物在一个干净，干燥的通畅的区域。线笼（高压灭菌）用干净的纸巾和调节水毯的温度升高到大约20至22℃。提供笼地板干燥和潮湿的粗粒自由采食。 注意:手术后护理的一个重要组成部分是观察动物和适当的干预的，根据需要，从麻醉和手术后的恢复过程中。监测的必要强度将随动物以及在即时麻醉恢复期间作为共可能是更大的mpared到后来的术后恢复。 </li><li>持续监控其监控经历了麻醉，直到他们完全康复的动物。动物必须能够保持胸骨无助和卧它必须出现平静和自由的痛苦，才可以无人看管。 </li><li>给予丁丙诺啡（0.1毫克/千克，BID）和酮洛芬（5毫克/公斤，SID），为期三天​​，都皮下。 </li><li>恢复从麻醉最少三天期间监测心血管功能和呼吸功能，体温，和手术后疼痛或不适。附加医疗可能需要，如止痛药和其他药物，并尽量减少脱水和电解质的丢失肠道液体给药。 </li><li>通过观察后肢运动功能评估移植手术的成功。移植的圆满成功涉及通畅的血液流向后肢和尾巴，这应该是阿尼姆的恢复时立即人，并完成恢复的后肢在第二天没有瘫痪</li></ol></li></ol> 3.组织收集注意:预确定终点的范围从3-60天，取决于分析所需的类型和抗原错配的性质。一般情况下，内膜增厚是整个完整的MHC不匹配的小鼠品系移植后第30天强劲。 <ol><li>麻醉小鼠用氯胺酮的腹膜内注射（100mg / kg的10毫克/毫升）和甲苯噻嗪（10毫克/千克; 1毫克/毫升）。按捏动物垫脚的脂肪部分评估麻醉深度。手术进行时，动物不会出现撤退反射。 </li><li>清洁手术部位水，用70％的酒精结束。 </li><li>发生在仰卧位动物的内衬吸水垫托盘。 </li><li>打开腹腔的大中线纵形切口。放置一个自固牵开，以暴露所述腔，下腔静脉的d为腹主动脉。 </li><li>轻轻分开来自相邻IVC使用一对医疗5号镊子的移植供体主动脉段。要特别小心，不要剥去移植的血管外膜。 </li><li>通过沿着胸椎一路胸廓入口两侧的肋切割暴露胸腔。反映前胸壁优揭露心包和安全的肋骨有一对止血钳。 </li><li>一旦心脏露出，插入地下25 5/8针（附连到注射器）进入左心室并用0.1ml肝素（100单位/毫升）的盐溶液冲洗。 </li><li>使用剪刀，取出右心房，以允许血液，肝素盐水和固定液灌注期间离开身体。灌注动物用5ml 4％多聚甲醛或直到流体运行明确。取出心脏，以确保安乐死。 </li><li>消费移植的血管段，沉浸在4％的对甲醛为不超过一小时以上。坐落在华侨城移植的血管（最佳切削温度）矩阵和闪光冻结。节下低温恒温器设定在8微米厚度的容器中。染色切片用苏木精 - 伊红和/或维尔赫夫面包车Gieson染色（面包车Gieson染色）弹性污点。 </li></ol>移植的4形态分析注意:TA的特征在于内膜增厚13。在这种模式下，内膜增厚和所得减少内腔的尺寸是反射免疫介导的血管损伤的严重程度。 Syngraft控制用于确定在没有同种异体免疫反应血管的基线特征。这些控制也允许发生作为外科手术的结果，动脉损伤的评价。应该有syngrafts没有内膜增厚。 <ol><li>要检查内膜增厚及管腔闭塞，内皮细胞内测量面积层，内弹力层（IEL）和弹性面包车Gieson染色染色动脉段与图像分析软件外弹性膜（鳗鲡）。下面的参数可被用于确定动脉变化的反射的TA。 <ol><li>测量绝对内膜面积:内部弹性层内区域 - 在内皮细胞层内的区域。这提供了内膜扩张，这是TA的标志的绝对测量。 </li><li>测量绝对内侧区:区域外弹性膜内 - 区域内弹性膜之内。这提供了内侧降解或生长的绝对测量，偶尔可发生的免疫介导的变化的血管壁的该区域的结果。 </li><li>测量总血管面积作为外部弹性层内的区域。这提供了血管重塑的测量涉及扩张或动脉的收缩，作为免疫学损伤的结果。 </li><li>测量内膜/介质:（区域的内弹性层内 - 的内皮细胞层内面积）/（所述外弹性膜内区域 - 内部弹性层内的区域）。这提供了内膜扩张的相对度量并标准化为在容器大小的差异。 </li><li>测量％管腔变窄:[（内弹性层内区域 - 在内皮细胞层内的区域）与内部弹性层/面积）] * 100。这提供了管腔闭塞的程度，这导致从内膜扩张和血管壁重塑（向内或向外）的组合的量度。 </li></ol></li></ol>

<ol>
	<li>Billingham, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Graft+coronary+disease:+the+lesions+and+the+patients.">Graft coronary disease: the lesions and the patients.</a> Transplant Proc. 21, 3665-3666 (1989).</li><li>Foegh, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+rejection--graft+arteriosclerosis.">Chronic rejection--graft arteriosclerosis.</a> Transplant Proc. 22, 119-122 (1990).</li><li>Libby, P., Pober, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+rejection.">Chronic rejection.</a> Immunity. 14, 387-397 (2001).</li><li>Tellides, G., Pober, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interferon-gamma+axis+in+graft+arteriosclerosis.">Interferon-gamma axis in graft arteriosclerosis.</a> Circulation research. 100, 622-632 (2007).</li><li>Johnson, D. E., Gao, S. Z., Schroeder, J. S., DeCampli, W. M., Billingham, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+spectrum+of+coronary+artery+pathologic+findings+in+human+cardiac+allografts.">The spectrum of coronary artery pathologic findings in human cardiac allografts.</a> The Journal of heart transplantation. 8, 349-359 (1989).</li><li>Gao, S. Z., Alderman, E. L., Schroeder, J. S., Silverman, J. F., Hunt, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+coronary+vascular+disease+in+the+heart+transplant+patient:+coronary+arteriographic+findings.">Accelerated coronary vascular disease in the heart transplant patient: coronary arteriographic findings.</a> Journal of the American College of Cardiology. 12, 334-340 (1988).</li><li>Mennander, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+rejection+in+rat+aortic+allografts.+An+experimental+model+for+transplant+arteriosclerosis.">Chronic rejection in rat aortic allografts. An experimental model for transplant arteriosclerosis.</a> Arterioscler Thromb. 11, 671-680 (1991).</li><li>Choy, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Granzymes+and+perforin+in+solid+organ+transplant+rejection.">Granzymes and perforin in solid organ transplant rejection.</a> Cell Death Differ. 17, 567-576 (2010).</li><li>Johnson, P., Carpenter, M., Hirsch, G., Lee, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recipient+cells+form+the+intimal+proliferative+lesion+in+the+rat+aortic+model+of+allograft+arteriosclerosis.">Recipient cells form the intimal proliferative lesion in the rat aortic model of allograft arteriosclerosis.</a> Am J Transplant. 2, 207-214 (2002).</li><li>Minami, E., Laflamme, M. A., Saffitz, J. E., Murry, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracardiac+progenitor+cells+repopulate+most+major+cell+types+in+the+transplanted+human+heart.">Extracardiac progenitor cells repopulate most major cell types in the transplanted human heart.</a> Circulation. 112, 2951-2958 (2005).</li><li>Yu, 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=AIP1+prevents+graft+arteriosclerosis+by+inhibiting+interferon-gamma-dependent+smooth+muscle+cell+proliferation+and+intimal+expansion.">AIP1 prevents graft arteriosclerosis by inhibiting interferon-gamma-dependent smooth muscle cell proliferation and intimal expansion.</a> Circ Res. 109, 418-427 (2011).</li><li>Miller, C., DeWitt, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+and+humoral+responses+to+major+and+minor+histocompatibility+antigens.">Cellular and humoral responses to major and minor histocompatibility antigens.</a> Transplant Proc. 5, 303-305 (1973).</li><li>Tsutsui, H., 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=Lumen+loss+in+transplant+coronary+artery+disease+is+a+biphasic+process+involving+early+intimal+thickening+and+late+constrictive+remodeling:+results+from+a+5-year+serial+intravascular+ultrasound+study.">Lumen loss in transplant coronary artery disease is a biphasic process involving early intimal thickening and late constrictive remodeling: results from a 5-year serial intravascular ultrasound study.</a> Circulation. 104, 653-657 (2001).</li><li>Rossum, A., Enns, W., Shi, P., MacEwan, G. E., Choy, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bim+regulates+allogeneic+immune+responses+and+transplant+arteriosclerosis+through+effects+on+T+cell+activation+and+death.">Bim regulates allogeneic immune responses and transplant arteriosclerosis through effects on T cell activation and death.</a> Arterioscler Thromb Vasc Biol. 34, 1290-1297 (2014).</li><li>Ho, M. K., Springer, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+distribution,+structural+characterization,+and+biosynthesis+of+Mac-3,+a+macrophage+surface+glycoprotein+exhibiting+molecular+weight+heterogeneity.">Tissue distribution, structural characterization, and biosynthesis of Mac-3, a macrophage surface glycoprotein exhibiting molecular weight heterogeneity.</a> J Biol Chem. 258, 636-642 (1983).</li><li>Inoue, T., Plieth, D., Venkov, C. D., Xu, C., Neilson, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibodies+against+macrophages+that+overlap+in+specificity+with+fibroblasts.">Antibodies against macrophages that overlap in specificity with fibroblasts.</a> Kidney international. 67, 2488-2493 (2005).</li><li>Shi, 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=Immunologic+basis+of+transplant-associated+arteriosclerosis.">Immunologic basis of transplant-associated arteriosclerosis.</a> Proc Natl Acad Sci U S A. 93, 4051-4056 (1996).</li><li>Skaro, A. I., 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=CD8++T+cells+mediate+aortic+allograft+vasculopathy+by+direct+killing+and+an+interferon-gamma-dependent+indirect+pathway.">CD8+ T cells mediate aortic allograft vasculopathy by direct killing and an interferon-gamma-dependent indirect pathway.</a> Cardiovasc Res. 65, 283-291 (2005).</li><li>Tellides, G., 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=Interferon-gamma+elicits+arteriosclerosis+in+the+absence+of+leukocytes.">Interferon-gamma elicits arteriosclerosis in the absence of leukocytes.</a> Nature. 403, 207-211 (2000).</li><li>Wang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interferon-gamma+induces+human+vascular+smooth+muscle+cell+proliferation+and+intimal+expansion+by+phosphatidylinositol+3-kinase+dependent+mammalian+target+of+rapamycin+raptor+complex+1+activation.">Interferon-gamma induces human vascular smooth muscle cell proliferation and intimal expansion by phosphatidylinositol 3-kinase dependent mammalian target of rapamycin raptor complex 1 activation.</a> Circ Res. 101, 560-569 (2007).</li><li>Soulez, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+perlecan+fragment+LG3+is+a+novel+regulator+of+obliterative+remodeling+associated+with+allograft+vascular+rejection.">The perlecan fragment LG3 is a novel regulator of obliterative remodeling associated with allograft vascular rejection.</a> Circ Res. 110, 94-104 (2012).</li><li>Choy, J. C., Kerjner, A., Wong, B. W., McManus, B. M., Granville, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perforin+mediates+endothelial+cell+death+and+resultant+transplant+vascular+disease+in+cardiac+allografts.">Perforin mediates endothelial cell death and resultant transplant vascular disease in cardiac allografts.</a> Am J Pathol. 165, 127-133 (2004).</li><li>Choy, J. 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=Granzyme+B+induces+endothelial+cell+apoptosis+and+contributes+to+the+development+of+transplant+vascular+disease.">Granzyme B induces endothelial cell apoptosis and contributes to the development of transplant vascular disease.</a> Am J Transplant. 5, 494-499 (2005).</li><li>Reis, E. D., 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=Dramatic+remodeling+of+advanced+atherosclerotic+plaques+of+the+apolipoprotein+E-deficient+mouse+in+a+novel+transplantation+model.">Dramatic remodeling of advanced atherosclerotic plaques of the apolipoprotein E-deficient mouse in a novel transplantation model.</a> Journal of vascular surgery. 34, 541-547 (2001).</li><li>Potteaux, 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=Suppressed+monocyte+recruitment+drives+macrophage+removal+from+atherosclerotic+plaques+of+Apoe-/-+mice+during+disease+regression.">Suppressed monocyte recruitment drives macrophage removal from atherosclerotic plaques of Apoe-/- mice during disease regression.</a> J Clin Invest. 121, 2025-2036 (2011).</li></ol>

我们已经描述了一个协议，用于主动脉插入接枝小鼠是用于研究免疫介导的血管性排斥和TA有用。该模型可用于研究的TA的原因以及新的治疗策略的发展。它已被用于在过去，建立在TA 14,17-21适应性免疫，细胞毒性T细胞应答，细胞因子介导的CD4 T细胞效应的反应，和抗体介导的移植物损伤的重要作用。在小鼠中动脉移植是困难的，因为该动物的明显小尺寸;然而，随着实践和尽职调查，成功?...

在过去的30多年，在免疫抑制药物的进步已经减少移植排斥反应所致的急性排斥反应，但慢性排斥反应仍然是一个主要挑战。慢性心脏移植排斥反应的主要表现是移植动脉硬化（TA）1,2。这种情况的特点由内膜增生和移植动脉血管舒缩功能障碍和发展为内皮细胞和平滑肌细胞的接受者免疫系统的免疫学定位的结果。移植物血管因承认外国肽主要组织相容性复合体（MHC）的具体目标是通过TA的发展，同时保留主船3只强调在移植动脉。为了保持这是TA不发生实验时，收件人是基因完全相同的供体或当收件人缺乏T细胞和B细胞4观察。免疫介导的血管损伤及dysfunctioÑ ​​引起内膜增厚和纤维化的发展，以及脂质和ECM蛋白的异常积累，在TA 5。内膜增厚趋于整个动脉树4-6同心。移植物丢失和死亡通常发生从同种异体移植物的动脉4的管腔闭塞导致渐进缺血的结果。 1991年，Mennander 等 7率先主动脉干预模型大鼠模型TA。一些研究小组已经适应随后此过程中使用的小鼠。在此模型中，同种异体移植主动脉段开发病灶具有特征媲美的TA在临床移植观察。这包括内膜增厚特征在于平滑肌细胞样细胞和受体白细胞7的积聚。在过去的二十年中该模型已被用于产生重要的洞察的血管损伤，排斥和TA的机制。它可以是我们编辑审查过程中动脉病理与免疫和血管反应的问题。抗原错配的影响选择适当解决这些问题的能力。 跨越完整MHC障碍移植允许已知参与器官移植排斥反应的免疫反应的全面评价。这包括直接CD4和CD8 T细胞识别和外国肽-MHC的靶向通过接枝衍生细胞呈现，间接的CD4（以及可能的CD8）的T细胞识别和由收件人抗原呈现接枝衍生同种异体抗原呈递细胞靶向和抗体同种抗原对血管细胞表面介导的8认可。但是，为了在完全MHC错配的实验损伤的血管反应可以比所观察到临床上不同。约翰逊等人 9表明，在整个完整的MHC错配屏障移植主动脉插入移植物，大部分内膜细胞受体的起源和不供体来源。这比在人类移植大多数内膜平滑肌细胞是供体来源9,10的观察不同。考虑到这一限制，已经开发了替代实验模型是涉及跨越次要组织相容性抗原错配接枝触发血管反应更接近于那些在临床移植11观察。虽然这些替代模式允许重要的结论可以做出关于驱动的TA，这引起血管排斥次要组织相容性抗原不匹配的移植物不完全重新投降那些发生在临床免疫进程的发展的血管反应。例如，次要组织相容性抗原是由接枝反应的抗体12差识别。鉴于上述考虑，要考虑的病理questi是非常重要的上时，选择在主动脉插入模型中使用的抗原不匹配的类型检查。在这里，我们描述了小鼠动脉介入移植了详细的方案。我们描述完整MHC错配的小鼠，但相同的协议被用于在其他抗原错配的小鼠品系接枝之间插入嫁接。

<table><tbody><tr><td>C57BL/6J (H-2b)</td><td>Jackson Laboratories, Bar Harbour ME</td><td>Strain# 000664</td><td></td></tr><tr><td>Balb/cBYJ</td><td>Jackson Laboratories, Bar Harbour ME</td><td>Strain# 001026</td><td></td></tr><tr><td>Ketamine Hydrochloride Injection USP 100 mg/ml</td><td>Ketalean</td><td>DIN 00612316</td><td></td></tr><tr><td></td><td></td><td></td><td></td></tr><tr><td>Xylazine Injection 20 mg/ml</td><td>Rompum</td><td>DIN 02169592</td><td></td></tr><tr><td>Ketoprofen Injection 100 mg/ml</td><td>Anafen</td><td>DIN 01938126</td><td></td></tr><tr><td>Butorphanol Tartrate&amp;nbsp;injection 10 mg/ml</td><td>Torbugesic</td><td>DIN 008450000</td><td></td></tr><tr><td>Buprenorphine Injection 0.3 mg/ml</td><td>Reckitt Benckiser</td><td>B.N. 5241</td><td></td></tr><tr><td>Atipamezole hydrochloride sterile injectable solution</td><td>Antisedan</td><td>DIN 02237744</td><td></td></tr><tr><td>Heparin Sodium Injection, USP, 1,000 units/ml</td><td>McKesson Distribution</td><td>DIN 02264315</td><td></td></tr><tr><td>Tears naturale ophthalmic ointment</td><td>Alcon</td><td>DIN 02082519</td><td></td></tr><tr><td>Stereomicroscope</td><td>Leica</td><td>M80</td><td></td></tr><tr><td>0.9% Sodium Chloride, sterile</td><td>Baxter Corporation</td><td></td><td></td></tr><tr><td>Lactated Ringer&amp;rsquo;s solution, sterile</td><td>Baxter Corporation</td><td></td><td></td></tr><tr><td>0.9% Sodium Chloride Injection, sterile, 10 ml</td><td>Baxter Corporation</td><td></td><td></td></tr><tr><td>Alcohol Prep Pads</td><td>Loris</td><td></td><td></td></tr><tr><td>Povidone Iodine</td><td>Betadine</td><td></td><td></td></tr><tr><td>Chlorohexidine Gluconate 4% w/v</td><td>Germi-Stat</td><td></td><td></td></tr><tr><td>Black Polyamide Monofilament</td><td>AROSurgical Instruments</td><td>T4A10Q07</td><td></td></tr><tr><td>Suture, 10-0 suture, 70 microns</td><td>Corporation</td><td></td><td></td></tr><tr><td>Blue monofilament suture 5-0, P3 needle</td><td>Ethicon</td><td>8698G</td><td></td></tr><tr><td>1 ml Syringe</td><td>BD</td><td>REF 309659</td><td></td></tr><tr><td>10 ml Syringe</td><td>BD</td><td>REF 309604</td><td></td></tr><tr><td>1cc TB insulin syringe with 28 G 1/2</td><td>BD</td><td>REF 309309</td><td></td></tr><tr><td>25 G 7/8, hypodermic needle</td><td>BD</td><td>REF 305124</td><td></td></tr><tr><td>27 G 1/2, hypodermic needle</td><td>BD</td><td>REF 305109</td><td></td></tr><tr><td>Colibri Retractor- 1.5 cm spread 4 cm</td><td>Fine Science Tools</td><td>17000-04</td><td></td></tr><tr><td>S&amp;amp;T CAF-4 Clip applying forceps, without lock</td><td>Fine Science Tools</td><td>00072-14</td><td></td></tr><tr><td>Supergrip forceps, S&amp;amp;T</td><td>Fine Science Tools</td><td>00632-11</td><td></td></tr><tr><td>Medical No.5 forceps</td><td>Fine Science Tools</td><td>11253-20</td><td></td></tr><tr><td>Lexer Baby Scissors</td><td>Fine Science Tools</td><td>14078-10</td><td></td></tr><tr><td>Micro Adson forceps serrated</td><td>Fine Science Tools</td><td>11018-12</td><td></td></tr><tr><td>Vannas-Tubingen microscissors</td><td>Fine Science Tools</td><td>15003-08</td><td></td></tr><tr><td>Micro clamps, b-1; 3.5mm x 1mm; 7mm length</td><td>Fine Science Tools</td><td>00396-01</td><td></td></tr><tr><td>Graefe-forceps, 10cm 1x2 teeth</td><td>Fine Science Tools</td><td>11054-10</td><td></td></tr><tr><td>Castroviejo with lock and tungsten jaws</td><td>Fine Science Tools</td><td>12565-14</td><td></td></tr><tr><td>Hot glass bead sterilizer</td><td>Inotech 250</td><td>IS-250 - Steri-250</td><td></td></tr><tr><td>Non-woven gauzes</td><td>Progene</td><td></td><td></td></tr><tr><td>Cotton Tipped Applicators</td><td>Puritan</td><td></td><td></td></tr><tr><td>Beard Trimmer</td><td>Wahl</td><td></td><td></td></tr><tr><td>Heating pad</td><td>Sunbeam</td><td></td><td></td></tr></tbody></table>

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.

在这种模式下，从一个只Balb / CYJ小鼠腹主动脉插入之C57B1 / 6受体的下腹主动脉。这使针对同种异体血管免疫性反应的一个综合评价。免疫介导的血管损伤在此模型中发起为最终结果的内膜增厚，管腔变窄和招募免疫细胞的血管修复反应（ 图1和2）。这些标准那么作为一个读出的同种免疫反应的严重程度，血管性排斥反应，和TA。该过程的成功可以通过移植动脉段的健壮内...

Watch this Scientific Journal Video about Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis at JoVE.com

Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis

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 ...

mouse-model-alloimmune-induced-vascular-rejection-transplant

Research

JoVE Journal

Medicine

8.4K 次观看.  Simon Fraser University. 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. 

视频: Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant vasculopathy (TVP).
The commonly used animal model for cardiovascular chronic rejection studies is the heterotopic heart transplant model performed in laboratory rodents. This model is used widely in experiments since Ono and Lindsey (3) published their technique. To analyze the findings in the blood vessels, the heart has to be sectioned and all vessels have to be measured.
Another method to investigate chronic rejection in cardiovascular questionings is the aortic transplant model (1, 2). In the orthotopic aortic transplant model, the aorta can easily be histologically evaluated (2). The PVG-to-ACI model is especially useful for CAV studies, since acute vascular rejection is not a major confounding factor and Cyclosporin A (CsA) treatment does not prevent the development of CAV, similar to what we find in the clinical setting (4). A7-day period of CsA is required in this model to prevent acute rejection and to achieve long-term survival with the development of TVP.
This model can also be used to investigate acute cellular rejection and media necrosis in xenogeneic models (5).

This video demonstrates the orthotopic aortic transplant model as a simple model to study the development of transplant vasculopathy (TVP) in rats.

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

原位主动脉瓣移植在小鼠血管疾病的研究

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

Mouse Models for Graft Arteriosclerosis

Graft arteriosclerois (GA), also called allograft vasculopathy, is a pathologic lesion that develops over months to years in transplanted organs characterized by diffuse, circumferential stenosis of the entire graft vascular tree. The most critical component of GA pathogenesis is the proliferation of smooth muscle-like cells within the intima. When a human coronary artery segment is interposed into the infra-renal aortae of immunodeficient mice, the intimas could be expand in response to adoptively transferred human T cells allogeneic to the artery donor or exogenous human IFN-&gamma; in the absence of human T cells. Interposition of a mouse aorta from one strain into another mouse strain recipient is limited as a model for chronic rejection in humans because the acute cell-mediated rejection response in this mouse model completely eliminates all donor-derived vascular cells from the graft within two-three weeks. We have recently developed two new mouse models to circumvent these problems. The first model involves interposition of a vessel segment from a male mouse into a female recipient of the same inbred strain (C57BL/6J). Graft rejection in this case is directed only against minor histocompatibility antigens encoded by the Y chromosome (present in the male but not the female) and the rejection response that ensues is sufficiently indolent to preserve donor-derived smooth muscle cells for several weeks. The second model involves interposing an artery segment from a wild type C57BL/6J mouse donor into a host mouse of the same strain and gender that lacks the receptor for IFN-&gamma; followed by administration of mouse IFN-&gamma; (delivered via infection of the mouse liver with an adenoviral vector. There is no rejection in this case as both donor and recipient mice are of the same strain and gender but donor smooth muscle cells proliferate in response to the cytokine while host-derived cells, lacking receptor for this cytokine, are unresponsive. By backcrossing additional genetic changes into the vessel donor, both models can be used to assess the effect of specific genes on GA progression. Here, we describe detailed protocols for our mouse GA models.

We describe protocols for our mouse graft arteriosclerois (GA) models which involve interposition of a mouse vessel segment into a recipient of the same inbred strain. By backcrossing additional genetic changes into the vessel donor, the model can assess the effect of specific genes on GA.

嫁接动脉硬化的小鼠模型

Graft arteriosclerois (GA),  also called allograft vasculopathy, is a pathologic lesion that develops over  months to years in transplanted organs ...

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 ...

In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients

The main concern in transplantation is to achieve specific tolerance through induction of regulatory cells. The understanding of tolerance mechanisms requires reliable models. Here, we describe models of tolerance to cardiac allograft in rat, induced by blockade of costimulation signals or by upregulation of immunoregulatory molecules through gene transfer. Each of these models allowed in vivo generation of regulatory cells such as regulatory T cells (Tregs), regulatory B cells (Bregs) or regulatory myeloid cells (RegMCs). In this manuscript, we describe two complementary protocols that have been used to identify and define in vitro and in vivo regulatory cell activity to determine their responsibility in tolerance induction and maintenance. First, an in vitro suppressive assay allowed rapid identification of cells with suppressive capacity on effector immune responses in a dose dependent manner, and can be used for further analysis such as cytokine measurement or cytotoxicity. Second, the adoptive transfer of cells from a tolerant treated recipient to a newly irradiated grafted recipient, highlighted the tolerogenic properties of these cells in controlling graft directed immune responses and/or converting new regulatory cells (termed infectious tolerance). These methods are not restricted to cells with known phenotypic markers and can be extended to any cell population. Furthermore, donor directed allospecificity of regulatory cells (an important goal in the field) can be assessed by using third party donor cells or graft either in vitro or in vivo. Finally, to determine the specific tolerogenic capacity of these regulatory cells, we provide protocols to assess the humoral anti-donor antibody responses and the capacity of the recipient to develop humoral responses against new or former known antigens. The models of tolerance described can be used to further characterize regulatory cells, to identify new biomarkers, and immunoregulatory molecules, and are adaptable to other transplantation models or autoimmune diseases in rodent or human.

In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients

Here, we present a protocol to induce tolerance in transplantation, and assess in vitro and in vivo the suppressive capacity of distinct cell subsets from the recipient and the immune status of the recipient toward donor or exogenous antigens.

体外和活体评估移植受体的 T、B 和髓细胞抑制活性和体液反应

The main concern in transplantation is to achieve specific tolerance through induction of regulatory cells. The understanding of tolerance mechanisms ...

免疫与感染

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 ...

Orthotopic Hind Limb Transplantation in the Mouse

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and translational research in the field of transplantation medicine. A vast array of reagents is available exclusively in this setting, including mono- and polyclonal antibodies for both diagnostic and interventional applications. In addition, a vast number of genotyped, inbred, transgenic, and knock out strains allow detailed investigation of the individual contributions of humoral and cellular components to the complex interplay of an immune response and make the mouse the gold standard for immunological research. 
Vascularized Composite Allotransplantation (VCA) delineates a novel field of transplantation using allografts to replace "like with like" in patients suffering traumatic or congenital tissue loss. This surgical methodological protocol shows the use of a non-suture cuff technique for super-microvascular anastomosis in an orthotopic mouse hind limb transplantation model. The model specifically allows for comparison between established paradigms in solid organ transplantation with a novel form of transplants consisting of various different tissue components. Uniquely, this model allows for the transplantation of a viable vascularized bone marrow compartment and niche that have the potential to exert a beneficial effect on the balance of immune acceptance and rejection. This technique provides a tool to investigate alloantigen recognition and allograft rejection and acceptance, as well as enables the pursuit of functional nerve regeneration studies to further advance this novel field of transplantation.

This novel model for orthotopic hind limb transplantation in the mouse, applying a non-suture cuff technique for super-microvascular anastomosis, provides a powerful tool for in&#160;vivo mechanistic immunological research related to vascularized composite allotransplantation (VCA).

原位后肢移植中的鼠标

In vivo animal model systems, and in particular mouse models, have evolved into powerful and versatile scientific tools indispensable to basic and ...

Modified Heterotopic Hindlimb Osteomyocutaneous Flap Model in the Rat for Translational Vascularized Composite Allotransplantation Research

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand and face transplants and, more recently, abdominal wall, uterus, and urogenital transplants. Functional outcomes have exceeded initial expectations, and most recipients enjoy an improved quality of life. However, as clinical experience accumulates, chronic rejection and complications from the immunosuppression must be addressed. In many cases where grafts have failed, the causative pathology has been ischemic vasculopathy. <span style="font-size: 11pt; line-height: 115%; font-family: Arial, sans-serif; color: rgb(41, 43, 49); background-image: initial; background-position: initial; background-size: initial; background-repeat: initial; background-attachment: initial; background-origin: initial; background-clip: initial;">The biological mechanisms of the acute and chronic rejection associated with VCA, especially ischemic vasculopathy, are important areas of research. However, due to the very small number of VCA patients, the evaluation of proposed mechanisms is better addressed in an experimental model. Multiple groups have used animal models to address some of the relevant unsolved questions in VCA rejection and vasculopathy. Several model designs involving a variety of species are described in the literature. Here we present a reproducible model of VCA heterotopic hindlimb osteomyocutaneous flap in the rat that can be utilized for translational VCA research. This model allows for the serial evaluation of the graft, including biopsies and different imaging modalities, while maintaining a low level of morbidity.

Vascularized composite allograft offers life-altering benefits to transplant recipients, but the biological causes of graft rejection and vasculopathy remain poorly understood. The rodent surgical model presented here offers a reproducible, clinically relevant model of transplantation, allowing researchers to evaluate rejection events and potential therapeutic strategies to prevent their occurrence.

改良异位异体下肢骨皮瓣模型在大鼠转化血管化复合同种异体移植研究中的应用

Vascularized composite allotransplantation (VCA) is a relatively new field in the reconstructive surgery. Clinical achievements in human VCA include hand ...

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. ...

A Rat Orthotopic Renal Transplantation Model for Renal Allograft Rejection

Renal allograft rejection limits the long-term survival of patients after renal transplantation. Rat orthotopic renal transplantation is an essential model to investigate the mechanism of renal allograft rejection in pre-clinical studies and could aid in the development of novel approaches to improve the long-term survival of renal allografts. Donor kidney implantation in rat orthotopic renal transplantation is commonly performed by end-to-side anastomosis to recipients' aorta and inferior vena cava. In this model, the donor's kidney was implanted using end-to-end anastomosis to the recipients' renal artery and renal vein. The donor's ureter was anastomosed to the recipient's bladder in an end-to-side 'tunnel' method. This model contributes to better healing of ureter-bladder anastomosis and increases the recipients' survival by avoiding interference with blood supply and venous reflux of the lower body. This model can be used to investigate the mechanisms of acute and chronic immune and pathologic rejection of renal allografts. Here, the study describes the detailed protocols of this orthotopic renal transplantation between rats.

The rat orthotopic renal transplantation model contributes to investigating the mechanism of renal allograft rejection. The current model increases the recipients' survival without interference with blood supply and venous reflux of the lower body using an end-to-end anastomosis of kidney implantation and an end-to-side "tunnel" method of ureter-bladder anastomosis.

一种用于肾同种异体排斥的大鼠原位肾移植模型

Renal allograft rejection limits the long-term survival of patients after renal transplantation. Rat orthotopic renal transplantation is an essential ...