Name: a pre-clinical porcine model of orthotopic heart transplantation
Uploaded: 2019-04-27T14:30:00
Description: Watch this Scientific Journal Video about A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation at JoVE.com

The authors have no acknowledgements.

The institutional animal care committee approved all experimental protocols and animals were treated following the &#8220;Guide for the Care and Use of Laboratory Animals&#8221; prepared by the Institute of Laboratory Animal Resources, National Research Council, 1996. Male Yorkshire pigs (40&#8211;50 kg) were used to perform the orthotopic heart transplants (animal size can vary according to the investigators discretion and experimental goals).
1. Donor Procedure
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	<li>Anesthetic inducti...

<ol>
	<li>Lund, L. H., Edwards, L. 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=The+Registry+of+the+International+Society+for+Heart+and+Lung+Transplantation:+Thirty-third+Adult+Heart+Transplantation+Report-2016;+Focus+Theme:+Primary+Diagnostic+Indications+for+Transplant.">The Registry of the International Society for Heart and Lung Transplantation: Thirty-third Adult Heart Transplantation Report-2016; Focus Theme: Primary Diagnostic Indications for Transplant.</a> The Journal of Heart and Lung Transplantation. 35 (10), 1158-1169 (2016).</li><li>Lund, L. H., Edwards, L. 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This manuscript describes a large-animal pre-clinical model of orthotopic heart transplantation. Various small animal models of heterotopic heart transplantation have been successfully used to study the effects of novel treatments to improve organ preservation and decrease ischemia-reperfusion injury11,12,13. Additionally, small animal models permit survival models and long-term follow-up to investigate the development of reject...

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure1. Although ischemic times of up to four hours are tolerated adequately, an ischemic time of greater than six hours is associated with inferior outcomes2. Primary graft dysfunction remains the principal cause of early morbidity and mortality following transplantation2,3. The causes of primary graft dysfunction are multifactorial and include the use of marginal organs, recipient pulmonary vascular disease, hyperacute rejection, and ischemia-reperfusion injury sustained at the time of transplantation3.
Multiple studies have investigated novel methods for donor heart preservation to reduce the incidence of primary graft dysfunction4,5,6,7. It is common practice to assess new techniques and treatments in murine models of ischemia-reperfusion injury or heterotopic heart transplantation. Additionally, small animal models permit survival models and long-term follow-up to investigate the development of rejection and cardiac allograft vasculopathy11,12,13. However, most of these strategies fail initial clinical pilot trials or never reach this stage. It is of paramount importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings.
The porcine heart is often considered the most anatomically similar to the human heart when using large-animal models. As such, it is an ideal platform to perform cardiac surgical research. However, there are several important factors to consider when using a porcine model. First, the tissue is typically described as fragile and friable, especially in the right atrium and the pulmonary artery, being prone to tears14. Additionally, the pig heart is considered sensitive to manipulation and prone to arrhythmias, which is why one should routinely administer an anti-arrythmetic to each animal at the beginning of the experiment. An important anatomical difference between the porcine model and clinical heart transplantation is the left hemiazygous vein in the swine which drains directly into the coronary sinus. This has to be ligated during the recipient procedure to avoid continuous bleeding. Finally, the porcine model is very sensitive to ischemia, but it is still appropriate for acute studies in heart transplantation15.
This manuscript describes a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies5,6,8,9.

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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.

This pre-clinical model has been used successfully since 19945,6,8,9. Table 1 demonstrates representative results from pressure-volume relationships and echocardiographic parameters taken at baseline, and 3 h post-transplantation in a set of 5 experiments. Although we see a decrease in myocadial contractility following transplantation, this was not statistically significant.</p...

Watch this Scientific Journal Video about A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation at JoVE.com

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

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.

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

It is important to evaluate new heart preservation strategies using clinically relevant large-animal model for a reliable clinical translation. The porcine heart model for research in cardiac surgery is a very useful one because human and porcine hearts share lots of similarities in terms of anatomy, physiology, even size, so it's an almost ideal platform for research. After opening the pericardium of an anesthetized donor pig, have an assistant carefully retract the right ventricular outflow tract inferiorly and the pulmonary artery to the left and carefully retract the aorta to the right to avoid direct lesions to the pulmonary artery.Use the cautery to dissect the aortal pulmonary space and clear the anterior aspect of the ascending aorta from the connective tissue. After carefully retracting the right ventricular outflow tract use a 4.0 PROLENE suture to place a purse string stitch on the proximal ascending aorta adventitia and secure the suture with a tourniquet. Insert the cardioplegia delivery cannula into the ascending aorta within the purse string suture and tighten the tourniquet to secure the cannula.Next open the left inferior pulmonary vein and the superior and inferior vena cavae to ensure adequate cardiac venting and place the aortic cross-clamp on the distal ascending aorta. Initiate cardioplegia infusion targeting an aortic root pressure of 80 to 100 millimeters of mercury. Apply a 0.9%sodium chloride ice slush to the thoracic cavity and over the organ.When the cardioplegia infusion is finished, proceed with the cardiectomy in a traditional manner, sectioning the aorta after the innominate artery and the pulmonary artery and at the bifurcation to ensure a sufficient length for the implant. To initiate a cardiopulmonary bypass in the recipient, first use Metzenbaum and right angle forceps to dissect between the superior vena cava and the innominate artery and the inferior vena cava and the pericardium of the anesthetized recipient animal. Individually encircle the superior and inferior vena cavae with a 0-silk tie and secure each tape with a tourniquet.Use a 4-0 PROLENE suture to place two concentric purse string sutures on the distal ascending aorta adventitia and place purse string sutures on the inferior and superior vena cavae at the level of the pericardial reflection. Secure each suture with a tourniquet and use the Seldinger technique to insert a 17-21 French arterial cannula into the aorta. Use a 3/8-3/8 connector to connect the cannula to the arterial line of the bypass circuit.Make a five-millimeter incision at the center of the purse string sutures placed on the vena cavae and dilate the incision with a small angled instrument. Using a 24-28 French right-angled, single-stage venous cannula, cannulate the superior and inferior vena cavae, directing the angle superiorly in the superior vena cava and inferiorly into the inferior vena cava, away from the heart. Tighten the tourniquet holding the purse string suture to secure the cannula, and use a 3/8-3/8 1/2-Y connector to connect the cannulas to the venous line of the bypass circuit.Then initiate the cardiopulmonary bypass. After cardiopulmonary bypass initiation, use a sharp dissection instrument and a right angle to encircle the left hemiazygos vein and ligate the vessel distally with an O-silk tie. Cross-clamp the recipient aorta proximally to the arterial cannula and snare both vena cavae with the previously placed O-silk ties.Compare the left atrial cuff sizes of the donor and recipient and modify each as necessary. Deliver the first cardioplegia dose to the donor heart using the previously placed cardioplegia cannula. Next use the standard biatrial anastomotic technique and an SH needle to place one 4-0 PROLENE suture at the junction between the left atrium and the right inferior margin of the inferior vena cava.Place another suture at 180 degrees from the first suture to connect the donor and recipient cuffs, then complete the posterior and anterior wall anastomoses from the superior suture to the inferior suture. For the right atrium, open the donor right atrium from the appendage towards the inferior vena cava, creating a donor cuff that matches the recipient cuff size, then, starting at the junction between the inferior vena cava and the right atrium, complete the interior wall anastomosis and then the lateral wall. For the pulmonary artery, trim the edges of both the recipient and donor pulmonary arteries to create matching sizes.Use a BB needle to place a 5-0 PROLENE suture at the left lateral wall, uniting the donor and recipient vessels, and another at the right lateral edge. Then complete the inferior wall anastomosis and the anterior wall anastomosis. For the aorta, trim the edges as just demonstrated and place a suture on the left lateral wall connecting the donor and recipient vessels.Then complete the inferior wall and the anterior wall anastomosis before initiating a 60-minute reperfusion on the cardiopulmonary bypass. For a functional assessment of the graft, place a 0-silk tie around the inferior vena cava and insert a pressure-volume conductance catheter into the left ventricle through a small apical ventriculotomy to permit continuous measurements of the left ventricular pressure-volume relations. Here representative results from pressure-volume relationships and echocardiographic parameters taken at baseline, and at three hours post-transplantation in a set of five experiments are shown.During steady-state assessments, volume-dependent parameters are recorded such as the maximum and minimum rate of developed pressure. Volume-independent parameters are obtained by intermittent occlusion of the inferior vena cava during which the volume of the left ventricle progressively decreases and the end-systolic and end-diastolic pressure-volume relationships are recorded. The pre-load recruitable stroke work data can be used to determine the relationship between the stroke work and the corresponding end-diastolic volume.Various other metabolic parameters can also be measured with this model. This procedure requires at least one trained surgeon and approximately five practice experiments to optimize the research protocol for each research group. The porcine heart is fragile, friable, and especially sensitive to manipulation and ischemia, thus being prone to tears.It is important to revise all anastomoses to ensure minimal to no bleeding. This protocol can help in the development of new myocardial preservation strategies to decrease ischemia-perfusion injury and to prevent primary graft failure.

a-pre-clinical-porcine-model-of-orthotopic-heart-transplantation

Research

JoVE Journal

Medicine

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.

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

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.

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

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.

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

Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are attractive druggable therapeutic targets. Here we describe an efficient four-step strategy for pre-clinical evaluation of tyrosine kinase inhibitors (TKIs) in the treatment of acute leukemia. Initially, western blot analysis is used to confirm target inhibition in cultured leukemia cells. Functional activity is then evaluated using clonogenic assays in methylcellulose or soft agar cultures. Experimental compounds that demonstrate activity in cell culture assays are evaluated in vivo using NOD-SCID-gamma (NSG) mice transplanted orthotopically with human leukemia cell lines. Initial in vivo pharmacodynamic studies evaluate target inhibition in leukemic blasts isolated from the bone marrow. This approach is used to determine the dose and schedule of administration required for effective target inhibition. Subsequent studies evaluate the efficacy of the TKIs in vivo using luciferase expressing leukemia cells, thereby allowing for non-invasive bioluminescent monitoring of leukemia burden and assessment of therapeutic response using an in vivo bioluminescence imaging system. This strategy has been effective for evaluation of TKIs in vitro and in vivo and can be applied for identification of molecularly-targeted agents with therapeutic potential or for direct comparison and prioritization of multiple compounds.

Receptor tyrosine kinases are ectopically expressed in many cancers and have been identified as therapeutic targets in acute leukemia. This manuscript describes an efficient strategy for pre-clinical evaluation of tyrosine kinase inhibitors for the treatment of acute leukemia.

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. 
Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. 
Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Pre-clinical Model of Cardiac Donation after Circulatory Death

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after circulatory death (DCD) is a solution to address this limited availability, but due to a period of prolonged warm ischemia and the risk of tissue injury, its routine use in cardiac transplantation is seldom seen. In this manuscript we provide a detailed protocol closely mimicking current clinical practices in the context of DCD with continuous monitoring of heart function, allowing for the evaluation of novel cardioprotective strategies and interventions to decrease ischemia-reperfusion injury.
In this model, the DCD protocol is initiated in anesthetized Lewis rats by stopping ventilation to induce circulatory death. When systolic blood pressure drops below 30 mmHg, the warm ischemic time is initiated. After a pre-set warm ischemic period, hearts are flushed with a normothermic cardioplegic solution, procured, and mounted onto a Langendorff ex vivo heart perfusion system. Following 10 min of initial reperfusion and stabilization, cardiac reconditioning is continuously evaluated for 60 min using intraventricular pressure monitoring. A heart injury is assessed by measuring cardiac troponin T and the infarct size is quantified by histological staining. The warm ischemic time can be modulated and tailored to develop the desired amount of structural and functional damage. This simple protocol allows for the evaluation of different cardioprotective conditioning strategies introduced at the moment of cardioplegia, initial reperfusion and/or during ex vivo perfusion. Findings obtained from this protocol can be reproduced in large models, facilitating clinical translation.

This protocol shows a simple and flexible approach for the evaluation of new conditioning agents or strategies to increase the feasibility of cardiac donation after circulatory death.

Cardiac transplantation demand is on the rise; nevertheless, organ availability is limited due to a paucity of suitable donors. Organ donation after ...

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.

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

Orthotopic Kidney Auto-Transplantation in a Porcine Model Using 24 Hours Organ Preservation And Continuous Telemetry

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for kidney transplantation (KT). Even though, the use of allografts from extended criteria donors (ECD) could partially ease the shortage of organ donors, ECD organs carry a potentially higher risk for inferior outcomes and postoperative complications. Dynamic organ preservation techniques, modulation of ischemia-reperfusion and preservation injury, and allograft therapies are in the spotlight of scientific interest in an effort to improve allograft utilization and patient outcomes in KT.
Preclinical animal experiments are playing an essential role in translational research, especially in the medical device and drug development. The major advantage of the porcine orthotopic auto-transplantation model over ex vivo or small animal studies lies within the surgical-anatomical and physiological similarities to the clinical setting. This allows the investigation of new therapeutic methods and techniques and ensures a facilitated clinical translation of the findings. This protocol provides a comprehensive and problem-oriented description of the porcine orthotopic kidney auto-transplantation model, using a preservation time of 24 hours and telemetry monitoring. The combination of sophisticated surgical techniques with highly standardized and state-of-the-art methods of anesthesia, animal housing, perioperative follow up, and monitoring ensure the reproducibility and success of this model.

Large animal models play an essential role in preclinical transplantation research. Due to its similarities to the clinical setup, the porcine model of orthotopic kidney auto-transplantation described in this article provides an excellent in vivo setting for the testing of organ preservation techniques and therapeutic interventions.

In the present era of organ transplantation with critical organ shortage, various strategies are employed to expand the pool of available allografts for ...