作者感谢她的组织学工作的克里斯蒂安Pahrmann。 资助宋佳Schrepfer已收到由德意志研究联合会（DFG）（SCHR992 / 3 1和SCHR992/4-1）的研究补助金。]

查尔斯河（Sandhofer WEG 7，D - 97633 Sulzfeld）均购自大鼠，体重约250 - 300G。 使用PVG大鼠作为捐助者，和作为接受者的ACI大鼠移植手术。 老鼠被安置在常规条件下，美联储标准大鼠议员和水广告libidum。 所有手术器械消毒后才能使用。 捐助者的准备: 与异氟醚麻醉大鼠（2,5-3％）使用感应室。 <ol><li>剃须的腹部和胸部的头发和地方大鼠在它的后面，并置于它的鼻子和嘴口罩，以保持麻醉。 </li><li>消毒腹部和胸部区域广泛使用普罗沃碘，下次使用80％乙醇，重复此步骤三次。 </li><li>检查捏后脚要确保足够anesthesized大鼠的反射。 </li><li>抽血，通过开放的腹部和腹主动脉切成一个洞的老鼠。 </li><li>小心打开胸腔，发现胸降主动脉。 </li><li>解剖胸主动脉周围组织如脂肪，神经和食管。 </li><li>撤回主动脉1.5厘米一块使用couter，未经组织损伤的主动脉。 </li><li>灌注主动脉移植冷无菌生理盐水彻底的移植，并存储在4 ° C。 </li></ol>收件人: 与异氟醚麻醉大鼠（2,5-3％）使用感应室。在手术过程中保持体温。 <ol><li>剃须腹部和应用眼膏，以防止在麻醉过程中干燥的眼睛。 </li><li>将大鼠在它的后面，并置于它的鼻子和嘴口罩，以保持麻醉。 </li><li>使用普罗沃碘，下次使用80％乙醇，重复此步骤三次消毒腹部。 </li><li>检查捏后脚要确保足够anesthesized大鼠的反射。 </li><li>执行中线腹部切口，分离两个步骤的皮肤和肌肉，打开腹部。 </li><li>放置在一个温暖的生理盐水滋润无粉手套的肠子。折叠的肠子周围的手套，以防止水分流失。 </li><li>取出的脂肪组织覆盖腹主动脉。 </li><li>解剖主动脉形成下腹地区分叉，注意不要造成损害血管的分支。 </li><li>如果必要的主动脉结扎分支。 </li><li>首先使用显微解剖主动脉肾部分钳，阻止血液的流动。 </li><li>下一步，放置第二钳接近主动脉分叉处。 </li><li>现在，血流停止，删除短段的主动脉。 </li><li>体主动脉瓣移植，在冰上一直保持在0.9％无菌生理盐水，缩短到足够的长度和位置的差距。 </li><li>连接到收件人主动脉捐助主动脉，执行运行缝线使用8-0普理灵缝合（爱惜康，日本东京，德国）。开始与颅结束对端吻合。 </li><li>当远端吻合完成后，小心地打开远端钳​​。 </li><li>如果没有出血远端钳开放后，小心地打开颅钳。缝合出血的情况下，再次关闭钳，找到出血，并使用一个单针止血。 </li><li>应当有一个可见的脉冲在远端的主动脉。 </li><li>下一步行动的肠子放回腹部。 </li><li>与预热的无菌生理盐水冲洗腹部。 </li><li>关闭的墙壁用6-0普理灵运行缝线（爱惜康，日本东京，德国）的腹部肌肉层。 </li><li>使用5-0普理灵（爱惜康，日本东京，德国），运行缝合关闭皮肤。 </li><li>虽然仍然是在麻醉大鼠，注入皮下4-5mg/kg卡洛芬。 </li><li>要为这种类型的程序提供足够的镇痛，安乃近添加到饮用水疼痛用药3天移植后（50毫克每100毫升安乃近）。 </li></ol>

<ol>
	<li>Deuse, T., Hoyt, G., Koyanagi, T., Robbins, R. C., Schrepfer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevention+and+Inhibition+But+Not+Reversion+of+Chronic+Allograft+Vasculopathy+by+FK778.">Prevention and Inhibition But Not Reversion of Chronic Allograft Vasculopathy by FK778.</a> Transplantation. 85, 870-877 (2008).</li><li>Mennander, A., Tiisala, S., Halttunen, J., Yilmaz, S., Paavonen, T., Hayry, P. <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+allografts.+An+experimental+model+for+transplant+arteriosclerosis.">Chronic rejection in rat allografts. An experimental model for transplant arteriosclerosis.</a> Arterioscler Thromb Vasc Biol. 11, 671-680 (1991).</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, 225-225 (1969).</li><li>Poston, R. S., Billingham, M., Hoyt, 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=Rapamycin+reverses+chronic+graft+vascular+disease+in+a+novel+cardiac+allograft+model.">Rapamycin reverses chronic graft vascular disease in a novel cardiac allograft model.</a> Circulation. 100, 67-100 (1999).</li><li>Schrepfer, S., Deuse, T., Koch-Nolte, F., Krieger, T., Haddad, M., Schäfer, H., Pelletier, M. P., Robbins, R. C., Reichenspurner, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FK778+in+Experimental+Xenotransplantation:+A+Detailed+Analysis+of+Drug+Efficacy.">FK778 in Experimental Xenotransplantation: A Detailed Analysis of Drug Efficacy.</a> J Heart Lung Transplant. 26, 70-77 (2007).</li></ol>

由于一些原因，我们发现主动脉原位移植模型，更准确的方式来探讨异位心脏移植模型相比，TVP发展: 主动脉移植模型的手术技术方面，体现的是一种方便可行的方法，只需要在月底到年底，吻合的成就，而异位心脏移植模型可能被视为一个更加复杂和容易出错的外科医生的技能挑战，需要在两个主要血管端至端侧吻合。 组织学标本的分析和测量大幅促进移植血管病变的...

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orthotopic aortic transplantation: a rat model to study the development of chronic vasculopathy

慢性排斥反应的研究模型是必不可少的调查期间移植血管病变（TVP）的发展的病理生物学和病理生理过程。 心血管慢性排斥反应的研究中常用的动物模型是在啮齿类实验动物进行异位心脏移植模型。这种模式是在实验中广泛使用，因为小野和Lindsey（3）发表了他们的的技术。要分析的结果，在血管，心，切片，所有船只都必须测量。 调查心血管questionings慢性排斥反应的另一种方法是主动脉移植模型（1，2）。主动脉主动脉原位移植模型，可以很容易地组织评估（2）。 PVG -预报模式是CAV的研究特别有用，因为急性血管性排斥反应的主要混杂因素和环孢素A（CsA）治疗并不妨碍CAV的发展，我们在临床上发现类似（4 ）。 A7天的环孢素A的时期，是需要在此模型中，以防止急性排斥反应，并实现长期生存与发展的TVP。 这种模式也可以用来研究急性细胞排斥反应和异种模型（5）媒体坏死。

Watch this Scientific Journal Video about Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy at JoVE.com

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

本视频演示了一个简单的模型来研究在大鼠体内的移植血管病变（TVP）的发展主动脉原位移植模型。

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 ...

orthotopic-aortic-transplantation-rat-model-to-study-development

Research

JoVE Journal

Medicine

12.1K 次观看.  University Hospital Hamburg. 本视频演示了一个简单的模型来研究在大鼠体内的移植血管病变（TVP）的发展主动脉原位移植模型。

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

Human Internal Mammary Artery (IMA) Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel stenting are crucial for translational approaches1,2.
The commonly used animal models include mice, rats, rabbits, and pigs3-5. However, the translation of these models into clinical settings remains difficult, since those biological processes are already studied in animal vessels but never performed before in human research models6,7. In this video we demonstrate a new humanized model to overcome this translational gap. The shown procedure is reproducible, easy, and fast to perform and is suitable to study the development of intimal hyperplasia and the applicability of diverse stents.
This video shows how to perform the stent technique in human vessels followed by transplantation into immunodeficient rats, and identifies the origin of proliferating cells as human.

Human Internal Mammary Artery IMA Transplantation and Stenting: A Human Model to Study the Development of In-Stent Restenosis

This video shows a model to study the development of intimal hyperplasia after stent deployment using a human vessel (IMA) in an immunodeficient rat model.

人乳内动脉（IMA）移植及内支架置入术:人体模型支架内再狭窄的研究开发

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel ...

科研

医学

Development of Obliterative Bronchiolitis in a Murine Model of Orthotopic Lung Transplantation

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and standardized not only for the study of allo-rejection but also between syngeneic strains for examining mechanisms of ischemia-reperfusion injury after lung transplantation. Although the application of the rat and other large animal model 2 contributed significantly to the elucidation of these studies, the scope of those investigations is limited by the scarcity of knockout and transgenic rats. Due to no effective therapies for obliterative bronchiolitis, the leading cause of death in lung transplant patients, there has been an intensive search for pre-clinical models that replicate obliterative bronchiolitis. The tracheal allograft model is the most widely used and may reproduce some of the histopathologic features of obliterative bronchiolitis 3. However, the lack of an intact vasculature with no connection to the recipient's conducting airways, and incomplete pathologic features of obliterative bronchiolitis limit the utility of this model 4. Unlike transplantation of other solid organs, vascularized mouse lung transplants have only recently been reported by Okazaki and colleagues for the first time in 2007 5. Applying the basic principles of the rat lung transplant, our lab initiated the obliterative bronchiolitis model using minor histoincompatible antigen murine orthotopic single-left lung transplants which allows the further study of obliterative bronchiolitis immunopathogenesis6.

Obliterative bronchiolitis is the key impediment to the long-term survival of lung transplant recipients and the lack of a robust preclinical model precludes examining obliterative bronchiolitis immunopathogenesis. Unlike other solid organ transplants, vascularized mouse lung transplantation has only recently been developed. Here we show our independently developed obliterative bronchiolitis model after murine orthotopic single-lung transplantation.

闭塞性细支气管炎在原位肺移植小鼠模型的发展

Orthotopic lung transplantation in rats was first reported by Asimacopoulos and colleagues in 1971 1. Currently, this method is well accepted and ...

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. ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

部分原位肝移植大鼠模型的肝动脉重建的外科手术

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

A Model of Chronic Nutrient Infusion in the Rat

Chronic exposure to excessive levels of nutrients is postulated to affect the function of several organs and tissues and to contribute to the development of the many complications associated with obesity and the metabolic syndrome, including type 2 diabetes. To study the mechanisms by which excessive levels of glucose and fatty acids affect the pancreatic beta-cell and the secretion of insulin, we have established a chronic nutrient infusion model in the rat. The procedure consists of catheterizing the right jugular vein and left carotid artery under general anesthesia; allowing a 7-day recuperation period; connecting the catheters to the pumps using a swivel and counterweight system that enables the animal to move freely in the cage; and infusing glucose and/or Intralipid (a soybean oil emulsion which generates a mixture of approximately 80% unsaturated/20% saturated fatty acids when infused with heparin) for 72 hr. This model offers several advantages, including the possibility to finely modulate the target levels of circulating glucose and fatty acids; the option to co-infuse pharmacological compounds; and the relatively short time frame as opposed to dietary models. It can be used to examine the mechanisms of nutrient-induced dysfunction in a variety of organs and to test the effectiveness of drugs in this context.

A protocol for chronic infusions of glucose and Intralipid in rats is described. This model can be used to study the impact of nutrient excess on organ function and physiological parameters.

在大鼠模型慢性营养输液

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each year. While several animal models exist for this procedure and mice are the species that is most commonly used. The reasons for using mice are the relative cost of using this species, the existence of many genetically defined strains that allow for the study of immune responses, and the existence of an extensive array of reagents that can be used to further define responses in this species. This model has been used to define factors in the cornea that are responsible for the relative immune privilege status of this tissue that enables corneal allografts to survive acute rejection in the absence of immunosuppressive therapy. It has also been used to define those factors that are most important in rejection of such allografts. Consequently, much of what we know concerning mechanisms of both corneal allograft acceptance and rejection are due to studies using a murine model of corneal transplantation. In addition to describing a model for acute corneal allograft rejection, we also present for the first time a model of late-term corneal allograft rejection.

Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

小鼠角膜移植模型，研究实体器官移植的最常见形式

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

正交心脏移植的临床前猪模型

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

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

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

大鼠原位肾移植: 一种新的简化的手术方法

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

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...