这项工作是由美国国立卫生研究院P30：奥布莱恩中心（DK 079337）的核心资源。

1。术前准备<ol><li>手术过程中，无论多么用心做，在显著压力的结果。为了尽量减少这种压力的影响，并最大限度地储备，动物应保持在动物饲养至少72小时之前使用10,11。 </li><li>必须是无菌的手术工具，纱布和棉签。这是没有必要使用无菌手套，提供的只是提示仪器触摸缝合或手术视野。 </li><li>用于止血，夹紧压力是非常重要的必要的最低限度。由于压力过大或粗糙处理而损坏的血管会导致血栓形成，导致在后，腿麻痹，肠缺血和死亡在24小时内。我们已经发现，使用与不超过2克/厘米2的压力下的夹具，消除了该问题。这些夹在表1中指出。钳的压力在规格夹由高品质的手术拉琴T制造商。我们发现，便宜夹或一次性夹普遍施加过大的压力。 </li><li>侧支血管的相对位置，以及腹主动脉和下腔静脉（IVC）的位置是应变依赖。您还可以找到小的小鼠个体之间的差异，特别是在侧支血管的位置。 </li></ol> 2。供体手术<ol><li>小鼠可以麻醉用异氟醚（1.5-2.5％的诱导，维持1.0％在空气中），或用戊巴比妥（55-65毫克/千克）。戊巴比妥规定的时间限制（1.5小时），但提供的外科医生选择旋转的动物更方便，如果需要的话。麻醉的时间可以延长至最多2小时到小肠或肝脏手术期间需要时，通过将2-3滴的麻醉。也可用于由训练有素的医生谁约氯胺酮/甲苯​​噻嗪（100毫克/千克体重7毫克/公斤二甲苯胺噻嗪，IP俗称“K仔”的氯胺酮及）Ň1至1.5小时内完成步骤，但它有一个窄的安全剂量范围。在美国，使用苯巴比妥已经成为更多的问题，在最近几年，由于供应短缺。至关重要的是，被监测麻醉深度，是一定的，它是足够的。脚趾掐反射是最好的指标，足够的深度，但也应注意呼吸速率和其他运动。 </li><li>麻醉诱导后，使用脱毛去除头发上腹部的凝胶或剃须刀。 </li><li>安装上的动物实验室磁带在扩展eagled的位置，以保持它的操作板采用。的金属板允许使用加热垫在它下面，在手术过程中的温度支持。我们手动控制的加热垫，这是一个消费级的单元，用于家庭使用的温度。温度被保持在35-38°之间。的金属的操作板也是易于消毒程序之间。 </li><li>使用杀菌剂如洗必泰消毒区域，随后用乙醇洗涤。重复此步骤三次，以确保充分消毒。这也将删除任何剩余的头发裁片，如果存在的话。 </li><li>用剪刀做一个切口，通过体壁，暴露腹腔脏器。 </li><li>使用棉签，内脏推到一边，露出腹部的大血管。的内脏放置一块纱布用温生理盐水弄湿保持湿润。 </li><li>使用钝性分离，非常仔细地解剖腹主动脉离周围组织。从下腔静脉解剖容器要非常小心。将一个或多个分支，如腰动脉，必须小心吊起并烧灼。解剖主动脉的部分应该是肾血管下方和上方的分叉进入股动脉的主动脉。这部分可以作为供体材料为许多的两个受助人，多少分支血管的供体动物的存在和大小。 </li><li>领带的缝合线的正下方的肾动脉和其他的正上方的分叉。用剪刀把主动脉，全含200 U / ml的肝素生理盐水冲洗移植。非常小心地保持一端的船只和滴生理盐水的解决方案，它贯穿于重力的船只。不要让气泡通过的船只。处理容器的端部，尽可能少尽可能其他的内膜可以损坏导致血栓形成购买。删除它从现场，并立即将其放置在冰冷的盐水的容器。允许供体动物的抽血。 </li></ol> 3。接收方操作<ol><li>麻醉动物和应用的脱毛胶或使用剃须刀去除头发上腹部。之前我们管理丁丙诺啡剂量在0.1毫克/千克建立镇痛皮下切口。 </li><li>中号指望。2-15动物实验室磁带的作业现场使用。动物必须装在一个温暖的操作阶段，这将有助于在操作过程中保持体温。 </li><li>庆大霉素眼药膏对眼睛的干燥，以防止他们。 </li><li>腹部，然后由乙醇洗消毒。重复此步骤三次，以确保充分消毒。 </li><li>再次检查是否有足够的麻醉，然后再进行下一个步骤。 </li><li>作出在两个阶段通过皮肤和体壁的正中切口，小心不要入射到组织下方。 </li><li>将吊具保持腹部切口开放。指向的螺杆组件朝向后肢端保持大量的钳位，满分的方式。 </li><li>一个正方形的无菌生理盐水纱布弄湿，并把它在肠道。随着你的手指轻轻地在纱布上，插入在肠道内的棉签，轻轻地反映到S的肠子IDE所以他们都坐在顶部的纱布。另一块纱布，把它的肠子，并用生理盐水湿。 </li><li>删除任何脂肪组织，，覆盖主动脉和下腔静脉（IVC）。要轻柔。的IVC是很脆弱的。 </li><li>轻轻地用钳子从下腔静脉解剖腹主动脉。清除的面积足够大，以提供空间两个夹具，在它们之间有足够的容器，这样，分割时，会有足够的接枝可缝合的容器。解剖时，寻找血管分支。如果必要的话，烧灼他们具有低温度cauterizer。 </li><li>将肾动脉和另一只以上的分叉血管钳的正下方。 </li><li>切主动脉。主动脉的端部通常会缩回，离开约5毫米的空间。检查止血。如果夹具的正常工作，只有少量的血逃脱后切。如果继续出血，检查的夹子，以确保没有干扰ING与他们同在。需要注意的是没有除去的主动脉。这仅仅是划分。 </li><li>用肝素盐水冲洗伤口两端（200 U /毫升），然后用棉签删除多余的解决方案。 </li><li>德移植到地方在每一端使用三个连续的缝合线。只是在每端一侧吻合缝合，然后填写在休息时进行连续缝合或间断缝合。七至八个不连续的缝线是足够的。如果您使用的是连续缝合，确保血管壁保持轻松，你不会导致吻合口狭窄。 </li><li>小心地取出钳颅端和查找内存泄漏的缝合线。少量渗水的确定提供约一分钟内停止。如果有更多的比，它是能够使用一个单一的线圈以止血。 </li><li>对主动脉的吻合口上方用湿棉签轻轻按下，然后删除第二个夹子。轻轻按下并释放了几次，检查显示主动脉专利。的接枝将立即灌注和脉冲应该是可见的。 </li><li>取出纱布覆盖的肠子，将它们放回原位。避免扭曲的肠道，并保持正常的解剖定位。 </li><li>关闭的肌肉层，用5-0薇乔。 </li><li>关闭的皮肤，使用5-0或6-0 prolene线。 </li><li>治疗小鼠皮下carprophen，在5毫克/公斤，以增加在终止前麻醉镇痛。 </li><li>给鼠标0.5-0.8毫升生理盐水皮下注射。 </li><li>将鼠标在加热笼恢复。在恢复过程中，请仔细监控，以确保恢复正常。手术后2-3小时，鼠标应该表现比较正常。在12小时后运，丁丙诺啡皮下2 mg / kg的镇痛治疗的动物。动物，在任何时候，如果弯腰驼背，制造噪音或表现出有限的运动范围，调查的问题。如果不能建立一个明确的原因和固定，应实施安乐死的动物，每个机构的协议。在24小时的动物应在收到另一个的carprophen剂量皮下注射5毫克/公斤。后肢麻痹​​表示失败的吻合或血栓。在这种情况下，动物应被安乐死。 </li></ol>

<ol>
	<li>Sun, 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=Improved+surgical+technique+for+the+establishment+of+a+murine+model+of+aortic+transplantation.">Improved surgical technique for the establishment of a murine model of aortic transplantation.</a> Microsurgery. 18, 368-371 (1998).</li><li>Dambrin, C., Calise, D., Pieraggi, M. T., Thiers, J. C., Thomsen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+aortic+transplantation+in+mice:+a+new+model+of+allograft+arteriosclerosis.">Orthotopic aortic transplantation in mice: a new model of allograft arteriosclerosis.</a> J. Heart Lung Transpl. 18, 946-951 (1999).</li><li>Calise, 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=Orthotopic+aortic+transplantation+in+rodents+by+the+sleeve+technique:+a+model+system+for+the+study+of+graft+vascular+disease.">Orthotopic aortic transplantation in rodents by the sleeve technique: a model system for the study of graft vascular disease.</a> Transpl. Proc. 33, 2369-2370 (2001).</li><li>Thomsen, 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=An+orthotopic+aortic+graft+mouse+model+to+study+the+immunopathology+of+chronic+vascular+rejection.">An orthotopic aortic graft mouse model to study the immunopathology of chronic vascular rejection.</a> Transpl. Proc. 34, 2833-2835 (2002).</li><li>Benza, R. L., George, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aortic+graft+transplantation+in+mice.">Aortic graft transplantation in mice.</a> J. Heart Lung Transpl. 21, 1319-1321 (2002).</li><li>Chereshnev, 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=Mouse+model+of+heterotopic+aortic+arch+transplantation.">Mouse model of heterotopic aortic arch transplantation.</a> The Journal of surgical research. 111, 171-176 (2003).</li><li>Cho, H. R., 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=Improved+surgical+technique+for+heterotopic+aortic+transplantation+in+mice.">Improved surgical technique for heterotopic aortic transplantation in mice.</a> J. Korean Med. Sci. 22, 12-15 (2007).</li><li>Zhong, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organ+transplantation+in+mice:+current+status+and+future+prospects.">Organ transplantation in mice: current status and future prospects.</a> Microsurgery. 19, 52-55 (1999).</li><li>George, J. F., Pinderski, L. J., Litovsky, S., Kirklin, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Of+mice+and+men:+mouse+models+and+the+molecular+mechanisms+of+post-transplant+coronary+artery+disease.">Of mice and men: mouse models and the molecular mechanisms of post-transplant coronary artery disease.</a> J. Heart Lung Transpl. 24, 2003-2014 (2005).</li><li>Gorska, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Principles+in+laboratory+animal+research+for+experimental+purposes.">Principles in laboratory animal research for experimental purposes.</a> Med Sci Monit. 6, 171-180 (2000).</li><li>Landi, M. S., Kreider, J. W., Lang, C. M., Bullock, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+shipping+on+the+immune+function+in+mice.">Effects of shipping on the immune function in mice.</a> Am. J. Vet. Res. 43, 1654-1657 (1982).</li><li>Sun, 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=Improved+surgical+technique+for+the+establishment+of+a+murine+model+of+aortic+transplantation.">Improved surgical technique for the establishment of a murine model of aortic transplantation.</a> Microsurgery. 18, 368-371 (1998).</li><li>Chen, B., 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=Carbon+monoxide+rescues+heme+oxygenase-1-deficient+mice+from+arterial+thrombosis+in+allogeneic+aortic+transplantation.">Carbon monoxide rescues heme oxygenase-1-deficient mice from arterial thrombosis in allogeneic aortic transplantation.</a> Am. J. Pathol. 175, 422-429 (2009).</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=Heart+transplantation+in+congenic+strains+of+mice.+Transpl.">Heart transplantation in congenic strains of mice. Transpl.</a> Proc. 5, 733-735 (1973).</li><li>Chang, E. 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=Vascular+anastomosis+using+controlled+phase+transitions+in+poloxamer+gels.">Vascular anastomosis using controlled phase transitions in poloxamer gels.</a> Nature medicine. 17, 1147-1152 (2011).</li></ol>

没有利益冲突的声明。

主动脉移植小鼠模型，因为小鼠很好定义immunogenetically 9,12,13，他们可以很容易地操纵，以改变它们的特定基因的表达，如果需要的话，提供了许多优点。正如在引进，血管外科在小鼠比大多数型号的船只的大小，因为是比较困难的。即使是伟大的动脉，如主动脉通常不超过100〜200微米内径操纵这些船只需要一个显着的技术和灵巧8,14。最常见的并发症是那些刚刚开始实施的系统观?...

<table border="1"><tbody><tr><td>项目</td><td>公司</td><td>产品编号</td></tr><tr><td>血管夹</td><td>精细科学工具</td><td> 00396-01（尺寸B-1） </td></tr><tr><td>杜蒙钳</td><td>精细科学工具</td><td> 11293-00 </td></tr><tr><td> 10-0针刺microsuture </td><td> AROSurgical </td><td> TK-107038 </td></tr><tr><td>直剪刀</td><td> Roboz手术仪器有限公司</td><td> RS-5620 </td></tr><tr><td>低温cauterizer </td><td>海狸Visitec国际</td><td> 8441000 </td></tr><tr><td>自护牵引器</td><td>世界精密仪器</td><td> 14240 </td></tr></tbody></table>

orthotopic aortic transplantation in mice for the study of vascular disease

在小鼠吻合血管手术通常被认为是困难的，高度依赖于个人的外科医生的技能。这在很大程度上是真实的，但也有一些重要的原则，可以减少这些程序的难度，并提高再现性。原位的主动脉移植是一个很好的学习过程，在这过程中这些原则，因为它涉及到两个终端到终端的吻合，但持续的成功需要有良好的缝合技术和处理的船只。这个过程开始于采购部门在收件人的原生主动脉的长度从供体动物，腹主动脉。采购主动脉收件人主动脉之间的分割端部，然后被放置到的地方使用的端至端吻合缝合。要成功实现这一目标，需要一个高集中度的好工具，稳健的手，和升值是多么容易鼠标就可以被损坏的血管，导致血栓形成。学习这些重要的原则是什么占据了大部分的初学者的时间学习显微小型啮齿动物时。在整个这个协议中，我们是指这些重要的点。该模型可用于研究血管性疾病中的各种不同的实验系统1-8。在这里示出的上下文中，它是最经常用于移植后的血管疾病，一个共同的实体器官移植的长期并发症内膜增生的发生在移植物的研究。该模型的主要优点是，它便于形态计量学分析和移植的血管位于邻接的内源性的容器，它可以作为一个附加的控制9。此处所示的技术是最经常使用小鼠体重18-25克。我们已经积累了我们的经验，使用C57BL/6J，BALB / CJ，和C3H/HeJ小鼠品系的。

图1示出了主动脉接枝。白色箭头表示的缝合线。专利接枝将显示一个可见的脉冲， 图2表示一个典型的实验中，受体存活56天的期间其次。一组由野生型（C57BL / 6×FVB）受体小鼠BALB / c小鼠主动脉移植。另一组“KO”，指定由缺乏表达血红素加氧酶1，从而导致血栓形成在24小时内移植的受惠人士半价优惠（C57BL / 6×FVB）。值得注意的是，这会导致所有收件人死亡作为图中所?...

Watch this Scientific Journal Video about Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease at JoVE.com

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

我们描述了一种技术，在该技术中的截面从一个鼠标的腹主动脉原位移植到刚好低于肾动脉在异体或同源的收件人。这种技术可能是有用的，大小均匀的大血管移植的研究中，被认为是有利的。

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. ...

orthotopic-aortic-transplantation-mice-for-study-vascular

Research

JoVE Journal

Medicine

10.8K 次观看.  The University of Alabama at Birmingham. 我们描述了一种技术，在该技术中的截面从一个鼠标的腹主动脉原位移植到刚好低于肾动脉在异体或同源的收件人。这种技术可能是有用的，大小均匀的大血管移植的研究中，被认为是有利的。

视频: Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

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 Small Bowel Transplantation in Rats

Small bowel transplantation has become an accepted clinical option for patients with short gut syndrome and failure of parenteral nutrition (irreversible intestinal failure). In specialized centers improved operative and managing strategies have led to excellent short- and intermediate term patient and graft survival while providing high quality of life 1,3. Unlike in the more common transplantation of other solid organs (i.e. heart, liver) many underlying mechanisms of graft function and immunologic alterations induced by intestinal transplantation are not entirely known6,7. Episodes of acute rejection, sepsis and chronic graft failure are the main obstacles still contributing to less favorable long term outcome and hindering a more widespread employment of the procedure despite a growing number of patients on home parenteral nutrition who would potentially benefit from such a transplant. The small intestine contains a large number of passenger leucocytes commonly referred to as part of the gut associated lymphoid system (GALT) this being part of the reason for the high immunogenity of the intestinal graft. The presence and close proximity of many commensals and pathogens in the gut explains the severity of sepsis episodes once graft mucosal integrity is compromised (for example by rejection). To advance the field of intestinal- and multiorgan transplantation more data generated from reliable and feasible animal models is needed. The model provided herein combines both reliability and feasibility once established in a standardized manner and can provide valuable insight in the underlying complex molecular, cellular and functional mechanisms that are triggered by intestinal transplantation. We have successfully used and refined the described procedure over more than 5 years in our laboratory 8-11. The JoVE video-based format is especially useful to demonstrate the complex procedure and avoid initial pitfalls for groups planning to establish an orthotopic rodent model investigating intestinal transplantation.

Small bowel transplantation has become an accepted treatment option for patients with irreversible intestinal failure. Our experimental model of orthotopic small bowel transplantation in rats serves as a reliable tool to address underlying immunologic and inflammatory processes that complicate intestinal transplantation.

大鼠原位小肠移植

Small  bowel transplantation has become an accepted clinical option for patients with  short gut syndrome and failure of parenteral nutrition ...

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

大鼠原位肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart failure. Here, we describe a detailed surgical procedure for creating pressure overload and cardiac hypertrophy in rats by constriction of the ascending aorta using a small metallic clip. After anesthesia, the trachea is intubated by inserting a cannula through a half way incision made between two cartilage rings of trachea. Then a skin incision is made at the level of the second intercostal space on the left chest wall and muscle layers are cleared to locate the ascending portion of aorta. The ascending aorta is constricted to 50&#8211;60% of its original diameter by application of a small sized titanium clip. Following aortic constriction, the second and third ribs are approximated with prolene sutures. The tracheal cannula is removed once spontaneous breathing was re-established. The animal is allowed to recover on the heating pad by gradually lowering anesthesia. The intensity of pressure overload created by constriction of the ascending aorta is determined by recording the pressure gradient using trans-thoracic two dimensional Doppler-echocardiography. Overall this protocol is useful to study the remodeling events and contractile properties of the heart during the gradual onset and progression from compensated cardiac hypertrophy to heart failure stage.

We describe a stepwise procedure for creating pressure overload and left ventricular hypertrophy in Wistar rats by constriction of the ascending aorta using a small metallic clip. This model is extensively used for studying remodeling changes during cardiac hypertrophy and for identifying and evaluating strategies for regression of such changes.

大鼠建立压力超负荷心肌肥厚模型在升主动脉缩窄

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

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 ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

主动脉钙化和炎症血管平滑肌细胞和成像的钙化

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

套筒技术在主动脉移植小鼠模型中的应用-教学视频

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Minimally Invasive Transverse Aortic Constriction in Mice

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard open-chest transverse aortic constriction (TAC). Although transverse aortic constriction is a highly functional method for the induction of high pressure in the left ventricle, it is a more difficult and lengthy procedure due to its use of artificial ventilation with tracheal intubation. TAC is oftentimes also less survivable, as the newer method, MTAC, neither requires the cutting of the ribs and intercostal muscles nor tracheal intubation with a ventilation setup. In MTAC, as opposed to a thoracotomy to access to the chest cavity, the aortic arch is reached through a midline incision in the anterior neck. The thyroid is pulled back to reveal the sternal notch. The sternum is subsequently cut down to the second rib level, and the aortic arch is reached simply by separating the connective tissues and thymus. From there, a suture can be wrapped around the arch and tied with a spacer, and then the sternal cut and skin can be closed. MTAC is a much faster and less invasive way to induce left ventricular hypertension and enables the possibility for high-throughput studies. The success of the constriction can be verified using high-frequency trans-thoracic echocardiography, particularly color Doppler and pulsed-wave Doppler, to determine the flow velocities of the aortic arch and left and right carotid arteries, the dimension of the blood vessels, and the left ventricular function and morphology. A successful constriction will also trigger significant histopathological changes, such as cardiac muscle cell hypertrophy with interstitial and perivascular fibrosis. Here, the procedure of MTAC is described, demonstrating how the resulting flow changes in the carotid arteries can be examined with echocardiography, gross morphology, and histopathological changes in the heart.

Minimally invasive transverse aortic constriction (MTAC) conserves the essentials of regular transverse aortic constriction (TAC) while eliminating the use of a ventilator with tracheal intubation. It proves to be a highly desirable method for high-throughput studies on left ventricular overload, particularly in translational studies.

微创横向主动脉缩窄小鼠

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

Induction of Nephrotic Syndrome in Mice by Retrobulbar Injection of Doxorubicin and Prevention of Volume Retention by Sustained Release Aprotinin

Nephrotic syndrome is the most extreme manifestation of proteinuric kidney disease and characterized by heavy proteinuria, hypoalbuminemia, and edema due to sodium retention and hyperlipidemia. To study the pathophysiology of this syndrome, rodent models have been developed based on the injection of toxic substances such as doxorubicin causing podocyte damage. In mice, only few strains are susceptible to this model. In wildtype 129S1/SvImJ mice, the administration of doxorubicin by rapid intravenous injection to the retrobulbar sinus induces experimental nephrotic syndrome that features all the symptoms of human disease including sodium retention and edema. After the onset of proteinuria, mice exhibit increased urinary serine protease activity that leads to the activation of the epithelial sodium channel (ENaC) and sodium retention. Pharmacological inhibition of urinary serine proteases by the treatment with sustained release aprotinin abrogates ENaC activation and prevents sodium retention. This model is ideal to study the pathophysiology of proteasuria, i.e., the excretion of active serine proteases that cause ENaC activation by the proteolysis of its &#947;-subunit. This can be regarded as the primary mechanism of ENaC activation and sodium retention in proteinuric kidney disease.

Here, we describe the induction of experimental nephrotic syndrome in 129S1/SvImJ mice by rapid retrobulbar injection of doxorubicin. We also treat nephrotic mice with sustained release pellets containing aprotinin to inhibit urinary serine protease activity and prevent sodium retention.

球注射阿霉素对小鼠肾病综合征的诱导作用及缓释抑肽酶预防容积保留率的研究

Nephrotic syndrome is the most extreme manifestation of proteinuric kidney disease and characterized by heavy proteinuria, hypoalbuminemia, and edema due ...

A Model of Reverse Vascular Remodeling in Pulmonary Hypertension Due to Left Heart Disease by Aortic Debanding in Rats

Pulmonary hypertension due to left heart disease (PH-LHD) is the most common form of PH, yet its pathophysiology is poorly characterized than pulmonary arterial hypertension (PAH). As a result, approved therapeutic interventions for the treatment or prevention of PH-LHD are missing. Medications used to treat PH in PAH patients are not recommended for treatment of PH-LHD, as reduced pulmonary vascular resistance (PVR) and increased pulmonary blood flow in the presence of increased left-sided filling pressures may cause left heart decompensation and pulmonary edema. New strategies need to be developed to reverse PH in LHD patients. In contrast to PAH, PH-LHD develops due to increased mechanical load caused by congestion of blood into the lung circulation during left heart failure. Clinically, mechanical unloading of the left ventricle (LV) by aortic valve replacement in aortic stenosis patients or by implantation of LV assist devices in end-stage heart failure patients normalizes not only pulmonary arterial and right ventricular (RV) pressures but also PVR, thus providing indirect evidence for reverse remodeling in the pulmonary vasculature. Using an established rat model of PH-LHD due to left heart failure triggered by pressure overload with subsequent development of PH, a model is developed to study the molecular and cellular mechanisms of this physiological reverse remodeling process. Specifically, an aortic debanding surgery was performed, which resulted in reverse remodeling of the LV myocardium and its unloading. In parallel, complete normalization of RV systolic pressure and significant but incomplete reversal of RV hypertrophy was detectable. This model may present a valuable tool to study the mechanisms of physiological reverse remodeling in the pulmonary circulation and the RV, aiming to develop therapeutic strategies for treating PH-LHD and other forms of PH.

The present protocol describes a surgical procedure to remove ascending-aortic banding in a rat model of pulmonary hypertension due to left heart disease. This technique studies endogenous mechanisms of reverse remodeling in the pulmonary circulation and the right heart, thus informing strategies to reverse pulmonary hypertension and/or right ventricular dysfunction.

大鼠主动脉切除术治疗左心病肺动脉高压反向血管重塑模型

Pulmonary hypertension due to left heart disease (PH-LHD) is the most common form of PH, yet its pathophysiology is poorly characterized than pulmonary ...