Name: Optimization of the Cuff Technique for Murine Heart Transplantation (Video) | JoVE Journal
Uploaded: 2020-06-26T13:00:00.000+00:00
Duration: 14 min 1 s
Description: Learn an optimized cuff technique for murine heart transplantation using an inner tube approach. This method facilitates vessel eversion and reduces operation time through collaborative surgical expertise.

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5 minutes

This work was supported by the Fujian Provincial Health Education Joint Research Project (WKJ2016-2-20), the National Natural Science Foundation of China (81771271 and 81800664), the National Key R&#38;D Program of China (2018YFA0108304) and the Education and Scientific Research Project for Young and Middle-aged Teachers in Fujian Province (JAT170714), Natural Science Foundation of Hunan Province of China (2019JJ50842) and Huxiang Young Talents of Hunan Province (2019RS2013).

Animals (BALB/c, C57BL/6, male, 8-12 weeks) are housed in a specific pathogen-free facility at the Xiamen University Laboratory Animal Center. C57BL/6 is used as recipient and BALB/c is used as donor. All the procedures are performed according to the Institutional Animal Care and Use Committee (IACUC) guidelines.
NOTE: A set of microsurgical instruments, including a micro scissor, micro straight forceps, micro curved forceps and micro needle holders, are necessary for the operation (Table and Materials, Figure 1B, C, D, E). One pair of single-use bulldog clamps (Figure 1F) is needed. Two cuffs for the external jugular vein and carotid artery are prepared by cutting the customized polyamide tubes with a No. 10 scalpel under a microscope. The diameter of the vein and artery cuff is 0.9 mm and 0.55 mm, respectively. In addition, the diameter of the inner tube for the corresponding vein cuff is 0.6 mm, and these of the inner tube for the corresponding artery cuff is 0.28&#160;mm (Figure 1G).
1. Recipient preparation
<ol>
	<li>Anesthetize the recipient mouse with pentobarbital (60 mg/kg, i.p). Use atraumatic mechanical clippers to remove the hair at the right lateral cervical region.</li>
	<li>Use a sterile cotton tip applicator to wipe the surgical area with iodine antiseptic followed by 70% ethanol.</li>
	<li>Place the mouse in the supine position on the operation platform. Cover the mouse with sterile gauze.</li>
	<li>Use an ophthalmic scissor to make a transverse incision from the lower one-third neck midline to the right shoulder-clavicle joint.</li>
	<li>Isolate the right external jugular vein with micro curved forceps to expose enough length, cut off the branches via electrocoagulation, and ligate the vessel at the distal end using a 6-0 silk suture.</li>
	<li>Clamp the external jugular vein proximally using a bulldog clamp and then transect the vein proximally to the ligature using a micro scissor.</li>
	<li>Wash the vessel lumen with 100 U/mL 0-4 &#176;C heparinized saline to remove any residual blood.</li>
	<li>Pull the external jugular vein through the vein cuff using micro straight forceps; insert the vein inner tube into the lumen as a stent, and evert the vessel wall over the cuff with micro straight forceps (Figure 2A).</li>
	<li>Fix the everted vessel endothelium at the proximal end of the cuff using a circumferential 8-0 silk suture (Figure 2B).</li>
	<li>Use micro straight forceps to withdraw the vein inner tube from the vein vessel.</li>
	<li>Perform blunt dissection with micro curved forceps to isolate the right carotid artery adjacent to the inner edge of the sternocleidomastoid.</li>
	<li>Clamp the right carotid artery proximally using a bulldog clamp, ligate the carotid artery distally using a 6-0 silk suture, and use a micro scissor to transect the carotid artery proximally to the ligature.</li>
	<li>Wash the carotid artery with 100 U/mL 0-4 &#176;C heparinized saline to remove any residual blood.</li>
	<li>Pass the carotid artery through the artery cuff and insert the artery inner tube into the artery vessel using micro straight forceps (Figure 2C).</li>
	<li>Evert the vessel over the cuff using micro straight forceps; fix the everted vessel endothelium using a circumferential 8-0 silk suture (Figure 2D).</li>
	<li>Withdraw the artery inner tube from the artery vessel with micro straight forceps. 
	NOTE: Preserve the submandibular gland of the recipient.</li>
</ol>
2. Donor preparation
<ol>
	<li>Anesthetize the donor mouse with pentobarbital (60 mg/kg, i.p). Use atraumatic mechanical clippers to remove the hair at the abdominal region.</li>
	<li>Place the mouse in the supine position on the operation platform. Cover the mouse with sterile gauze.</li>
	<li>Use a sterile cotton tip applicator to wipe the surgical area with iodine antiseptic followed by 70% ethanol.</li>
	<li>Make an abdominal midline incision with an ophthalmic scissor and expose the abdominal cavity.</li>
	<li>Use micro curved forceps to expose the inferior vena cava, and then intravenously inject 200 &#956;L of 100 U/mL 0-4 &#176;C heparinized saline per 20 g of bodyweight through the inferior vena cava.</li>
	<li>Perform thoracotomy with ophthalmic scissors, cut off the ribs through the bilateral midaxillary line incisions, flip the anterior chest wall outwards to expose the thoracic cavity.</li>
	<li>Excise the thymus with micro curved forceps.</li>
	<li>Expose the aorta, and then perfuse 200 &#181;L of 100 U/mL 0-4 &#176;C heparinized saline to the coronary artery through the aortic arch. 
	NOTE: Avoid perfusing any gas bubbles into the donor heart.</li>
	<li>Use a micro scissor to transect the ascending aorta at the beginning of the aortic arch.</li>
	<li>Transect the pulmonary artery at the beginning of the two main branches with a micro scissor.</li>
	<li>Ligate the superior vena cava and inferior vena cava proximally using a 6-0 silk suture and use a micro scissor to transect vein distally to the ligature.</li>
	<li>Ligate the pulmonary veins together, circumferentially, using a single 6-0 silk suture, and cut off the vein branches distally to the ligature using a micro scissor.</li>
	<li>Remove the heart graft from the surrounding soft tissues; preserve it in 0-4 &#176;C heparinized saline.</li>
</ol>
3. Heart implantation
<ol>
	<li>Place the donor heart upside down into the right neck region of the recipient.</li>
	<li>Input the pulmonary artery of the donor heart to a 6-0 silk loop with micro straight forceps.</li>
	<li>Wrap the vessel lumen around the vein cuff, and then tighten the 6-0 silk suture loops around the cuff to band the vessel joint.</li>
	<li>Perform anastomosis of the aorta of the graft and the artery cuff by following the steps described in step 3.2.</li>
	<li>Release the clamped jugular vein followed by the clamped jugular artery. Keep the vessel joint untwisted and ensure that the blood flow is unobstructed. 
	NOTE: The sinus rhythm returning to more than 200 times within 1 min is considered normal.</li>
	<li>Moisten the donor heart using warm (37 &#176;C) saline and inspect whether the graft is bleeding. Set the pulsing heart graft into the subcutaneous space, and then suture the incision.</li>
</ol>
4. Postoperative Care and Graft Assessment
<ol>
	<li>Record the time to normal sinus rhythm and the preservation of normal sinus rhythm for at least 5 minutes after clamp release to monitor the post-operative graft function.</li>
	<li>Place the recipient alone on a warm blanket until the recipient wakes up from anesthesia. Administer buprenorphine analgesia, 0.05 mg/kg, s.c, at the end of the surgery and every 12 hours for 72 hours post-surgery.</li>
	<li>Record the weight and postoperative recovery status of the recipient daily. In case of &#62;15% weight loss relative to that on the surgery date, hemiplegic paralysis, or infection, euthanize the recipient via terminal isoflurane inhalation21.</li>
	<li>Monitor graft survival by palpation daily. The surgery is considered to be successful if the murine allograft survives for &#62;72 hours. Grade the graft function, as previously reported22: Scale 3 - vigorously pulsate and frequency; Scale 2 - less pulsate; Scale 1- fibrillation and imminent rejection; or Scale - 0, loss of heart beat and complete rejection.</li>
</ol>

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<li>Chen, Z. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+technique+of+cervical+heterotopic+heart+transplantation+in+mice%5BTitle%5D)+AND+%22Transplantation%22%5BJournal%5D)">A technique of cervical heterotopic heart transplantation in mice.</a> Transplantation. 52 (6), 1099-1101 (1991).</li>
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<li>Chen, H., Zhang, Y., Zheng, D., Praseedom, R. K., Dong, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Orthotopic+kidney+transplantation+in+mice%3A+technique+using+cuff+for+renal+vein+anastomosis%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Orthotopic kidney transplantation in mice: technique using cuff for renal vein anastomosis.</a> PLoS One. 8 (10), 77278(2013).</li>
<li>Miller, M. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Spontaneous+restoration+of+transplantation+tolerance+after+acute+rejection%5BTitle%5D)+AND+%22Nature+Communications%22%5BJournal%5D)">Spontaneous restoration of transplantation tolerance after acute rejection.</a> Nature Communications. 6, 7566(2015).</li>
<li>Lin, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Overexpression+of+Jagged-1+combined+with+blockade+of+CD40+pathway+prolongs+allograft+survival%5BTitle%5D)+AND+%22Immunology+and+Cell+Biology%22%5BJournal%5D)">Overexpression of Jagged-1 combined with blockade of CD40 pathway prolongs allograft survival.</a> Immunology and Cell Biology. 93 (2), 213-217 (2015).</li>
<li>Xie, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Combined+costimulation+blockade+inhibits+accelerated+rejection+mediated+by+alloantigen-primed+memory+T+cells+in+mice%5BTitle%5D)+AND+%22Immunological+Investigations%22%5BJournal%5D)">Combined costimulation blockade inhibits accelerated rejection mediated by alloantigen-primed memory T cells in mice.</a> Immunological Investigations. 38 (7), 639-651 (2009).</li>
<li>Shao, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Combination+of+monoclonal+antibodies+with+DST+inhibits+accelerated+rejection+mediated+by+memory+T+cells+to+induce+long-lived+heart+allograft+acceptance+in+mice%5BTitle%5D)+AND+%22Immunology+Letters%22%5BJournal%5D)">Combination of monoclonal antibodies with DST inhibits accelerated rejection mediated by memory T cells to induce long-lived heart allograft acceptance in mice.</a> Immunology Letters. 138 (2), 122-128 (2011).</li>
<li>Dai, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Blockade+of+CD27%2FCD70+pathway+to+reduce+the+generation+of+memory+T+cells+and+markedly+prolong+the+survival+of+heart+allografts+in+presensitized+mice%5BTitle%5D)+AND+%22Transplant+Immunology%22%5BJournal%5D)">Blockade of CD27/CD70 pathway to reduce the generation of memory T cells and markedly prolong the survival of heart allografts in presensitized mice.</a> Transplant Immunology. 24 (4), 195-202 (2011).</li>
<li>Yan, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inhibition+of+accelerated+rejection+mediated+by+alloreactive+CD4%28%2B%29+memory+T+cells+and+prolonged+allograft+survival+by+arsenic+trioxide%5BTitle%5D)+AND+%22Immunological+Investigations%22%5BJournal%5D)">Inhibition of accelerated rejection mediated by alloreactive CD4(+) memory T cells and prolonged allograft survival by arsenic trioxide.</a> Immunological Investigations. 42 (5), 438-454 (2013).</li>
<li>Yan, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inhibiting+accelerated+rejection+mediated+by+alloreactive+CD4%28%2B%29+memory+T+cells+and+prolonging+allograft+survival+by+1alpha%2C25-dihydroxyvitamin+D%283%29+in+nude+mice%5BTitle%5D)+AND+%22Immunology+Letters%22%5BJournal%5D)">Inhibiting accelerated rejection mediated by alloreactive CD4(+) memory T cells and prolonging allograft survival by 1alpha,25-dihydroxyvitamin D(3) in nude mice.</a> Immunology Letters. 149 (1-2), 54-61 (2013).</li>
<li>Lin, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Arsenic+trioxide+is+a+novel+agent+for+combination+therapy+to+prolong+heart+allograft+survival+in+allo-primed+T+cells+transferred+mice%5BTitle%5D)+AND+%22Transplant+Immunology%22%5BJournal%5D)">Arsenic trioxide is a novel agent for combination therapy to prolong heart allograft survival in allo-primed T cells transferred mice.</a> Transplant Immunology. 25 (4), 194-201 (2011).</li>
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<li>Li, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+highly+reproducible+cervical+cuff+technique+for+rat-to-mouse+heterotopic+heart+xenotransplantation%5BTitle%5D)+AND+%22Xenotransplantation%22%5BJournal%5D)">A highly reproducible cervical cuff technique for rat-to-mouse heterotopic heart xenotransplantation.</a> Xenotransplantation. , (2017).</li>
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<li>Blanchard, J. M., Pollak, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Techniques+for+perfusion+and+storage+of+heterotopic+heart+transplants+in+mice%5BTitle%5D)+AND+%22Microsurgery%22%5BJournal%5D)">Techniques for perfusion and storage of heterotopic heart transplants in mice.</a> Microsurgery. 6 (3), 169-174 (1985).</li>
<li>Felix, N. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((H2-DMalpha%28-%2F-%29+mice+show+the+importance+of+major+histocompatibility+complex-bound+peptide+in+cardiac+allograft+rejection%5BTitle%5D)+AND+%22Journal+of+Experimental+Medicine%22%5BJournal%5D)">H2-DMalpha(-/-) mice show the importance of major histocompatibility complex-bound peptide in cardiac allograft rejection.</a> Journal of Experimental Medicine. 192 (1), 31-40 (2000).</li>
<li>Tomita, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Improved+technique+of+heterotopic+cervical+heart+transplantation+in+mice%5BTitle%5D)+AND+%22Transplantation%22%5BJournal%5D)">Improved technique of heterotopic cervical heart transplantation in mice.</a> Transplantation. 64 (11), 1598-1601 (1997).</li>
<li>Niimi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+technique+for+heterotopic+cardiac+transplantation+in+mice%3A+experience+of+3000+operations+by+one+surgeon%5BTitle%5D)+AND+%22Journal+of+Heart+and+Lung+Transplantation%22%5BJournal%5D)">The technique for heterotopic cardiac transplantation in mice: experience of 3000 operations by one surgeon.</a> Journal of Heart and Lung Transplantation. 20 (10), 1123-1128 (2001).</li>
<li>Wang, K., Zhang, N., Li, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Improved+technique+of+mouse+heterotopic+heart+graft+retransplantation%5BTitle%5D)+AND+%22Microsurgery%22%5BJournal%5D)">Improved technique of mouse heterotopic heart graft retransplantation.</a> Microsurgery. 26 (3), 200-202 (2006).</li>
<li>Plenter, R. J., Grazia, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Murine+heterotopic+heart+transplant+technique%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Murine heterotopic heart transplant technique.</a> Journal of Visualized Experiments. (89), (2014).</li>
<li>Ratschiller, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Heterotopic+Cervical+Heart+Transplantation+in+Mice%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiments%22%5BJournal%5D)">Heterotopic Cervical Heart Transplantation in Mice.</a> Journal of Visualized Experiments. (102), e52907(2015).</li>
<li>Zhou, Y., Gu, X., Xiang, J., Qian, S., Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+comparative+study+on+suture+versus+cuff+anastomosis+in+mouse+cervical+cardiac+transplant%5BTitle%5D)+AND+%22Experimental+and+Clinical+Transplantation%22%5BJournal%5D)">A comparative study on suture versus cuff anastomosis in mouse cervical cardiac transplant.</a> Experimental and Clinical Transplantation. 8 (3), 245-249 (2010).</li>
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</ol>

The authors have nothing to disclose.

Mouse heart transplants models are important tools for transplant immunology research, as tools and materials for evaluating the immune mechanisms of this model and a large number of gene-modified mice are available. However, microsurgical technical challenges, such as vessels suture and eversion, have limited its widespread use. In the present study, we have investigated certain core technical challenges of murine heart transplantation and have obtained good outcomes. A critical step of the protocol the insertion of an ...

The establishment of mouse heterotopic heart transplantation through end-to-end anastomosis within the abdomen in 1973 was a major milestone in basic transplant immunology research1. This model provided an important and valid tool for analyzing the mechanisms of ischemia reperfusion injury2, immunological rejection, and tolerance3,4. However, the complex and time-consuming nature of the surgery as well as the potential for infections can result in severe perioperative abdominal adhesions and inflammatory reactions, resulting in a low efficiency for heterotopic heart transplantation model.
The cervical heterotopic heart transplantation technique was first described by Chen in 19915. In this model, the recipient&#8217;s external jugular vein is anastomosed to the pulmonary artery of the graft and the carotid artery is anastomosed to the ascending aorta. The main advantages of this method are the convenience of monitoring and the reducing of trauma to the recipient. In the same year, Matsuura described an improved technique, in which the end of the external jugular vein and carotid artery were everted over a Teflon cuff and fixed with a circumferential silk ligature6. Some researchers also fixed the cuff to the right pulmonary artery in the donor heart before inserting the cuff into the external jugular vein of the recipient7. Thus far, the cuff technique has been widely applied in various vascular pedicle transplant models, including those for lung8, liver9, and renal10 transplantation.
To date, there are several difficulties associated with the cuff technique. For example, the carotid artery is difficult to evert over the cuff due to the additional elasticity, resulting in the tissue flipping backwards. Hence, additional practice and a microsurgical dilator may be required to complete this step. In addition, the cervical vessel preparation can take up to 25 minutes.
To resolve these issues, we introduce the inner tube technique, which is based on the cuff technique and includes fixing the cuff on the external jugular vein and carotid artery using an inner tube to helps with the eversion of the vessel wall. In addition, with simple training, the recipient preparation is reduced to 15.5 minutes. This technique reduces the complexity of the operation and does not require additional practice or the use of a vascular dilator. It can be applied in all transplantation immune research, especially for verifying third-party immune tolerance during which the recipient receives two cardiac allografts, one within the abdomen and the other in the neck11. We also recommend co-operation between two skilled surgeons to establish this model, with one surgeon preparing the recipient animal and the other harvesting and implanting the donor heart. Such collaboration can shorten the operation time to 25 minutes. Using this optimized procedure, we have established syngeneic, allogeneic12,13,14,15,16,17,18,19, and xenogeneic mouse heart transplantation models20.
The rationale for the development of the inner tube technique was to reduce the operation time for the establishment of a mouse heart transplant model with a high success rate. Optimization of the cervical heart transplantation model facilitates the acquisition of high success rates in a short period of surgery time compared to the traditional suture and cuff technique21. Further, the cooperation model can further reduce the warm ischemic time of the donor heart compared to surgeries performed with a single operator.

<table><tbody><tr><td>Artery cuff</td><td>Self-made</td><td></td><td>Polyamide tube. diameter: 0.55 mm,length: 1.0 mm</td></tr><tr><td>Artery inner tube</td><td>Self-made</td><td></td><td>Polyamide tube. Diameter: 0.28mm</td></tr><tr><td>Micro curved forceps</td><td>Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory</td><td>WA3050</td><td>1/8 arc, 0.3-mm tip without a hook</td></tr><tr><td>Micro needle holders</td><td>Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory</td><td>WA2050</td><td>0.2-mm tip</td></tr><tr><td>Micro scissors</td><td>Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory</td><td>WA1050</td><td>Straight, blade length: 10 mm</td></tr><tr><td>Micro straight forceps</td><td>Shanghai Medical Instruments (Group) Ltd., Corp. Surgical Instruments Factory</td><td>WA3060</td><td>0.15-mm tip without a hook</td></tr><tr><td>Scanlan Vascu-Statt Bulldog Clamps</td><td>Scanlan International Inc</td><td>1001-531</td><td>Clamping pressure 20&amp;ndash;25 grams</td></tr><tr><td>Vein cuff</td><td>Self-made</td><td></td><td>Polyamide tube. diameter: 0.9 mm,length: 1.2 mm</td></tr><tr><td>Vein inner tube</td><td>Self-made</td><td></td><td>Polyamide tube. Diameter: 0.6 mm</td></tr></tbody></table>

optimization of the cuff technique for murine heart transplantation

Optimization of the Cuff Technique for Murine Heart Transplantation

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

Surgical Operation Time
After training, a skilled surgeon can successfully perform the operation within 35 minutes using the inner tube technique, wherein approximately 15.5 minutes are required for recipient preparation, 10.9 minutes are required for donor preparation, and 4.4 minutes are required for donor heart anastomoses. The cold and warm ischemia time (from donor preparation to heart implantation) is reduced to 15.3 minutes compared to the operation using the cuff techn...

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

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

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Research

JoVE Journal

Immunology and Infection

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach. Herein, we provide additional details of this technique to supplement the video.

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

The manuscript describes the steps required to perform the heterotopic heart transplant in the mouse.

Murine Heterotopic Heart Transplant Technique

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

Heterotopic Cervical Heart Transplantation in Mice

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.

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

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

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model

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

Mouse Heterotopic Cervical Cardiac Transplantation Utilizing Vascular Cuffs

This protocol provides technical information for vessel reconstruction using the cuff technique in mouse orthotopic liver transplantation.

Vascular Reconstruction with the Cuff Technique in Mouse Orthotopic Liver Transplantation

This protocol presents a cuff-free, suture-based cervical heterotopic heart transplantation technique in mice, designed to enable the reliable placement of a second heterotopic cardiac graft for evaluating donor-specific immune tolerance under physiological conditions with minimal interference from foreign material.

Cuff-Free Suture-Based Cervical Model for Secondary Heterotopic Heart Transplantation to Evaluate Donor-Specific Immune Tolerance in Mice

This protocol presents a simplified, end-to-side suture anastomosis technique for murine cervical heart transplantation, providing a robust and reproducible model for studying transplant immunology.

Cervical Heterotopic Mouse Heart Transplantation using End-to-side Anastomosis Technique to Study Transplant Immunology

In this video, we demonstrate the procedure of CD40-activation and expansion of murine B cells from splenocytes of C57BL/6 mice, which can be used as a model antigen-presenting cell (APC) to study induction of immunity.

Murine Model of CD40-activation of B cells

Learn an optimized cuff technique for murine heart transplantation using an inner tube approach. This method facilitates vessel eversion and reduces operation time through collaborative surgical expertise.

Optimization of the Cuff Technique for Murine Heart Transplantation

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