This work was funded by a generous gift from the Bombeck Family Estate.

Lewis (RT11) and Dark Agouti (DA) (RT1Aa) rats were purchased from commercial vendors (see the Table of Materials). These fully MHC-mismatched strains are often used to study acute renal allograft rejection. All animals were housed and maintained according to the National Institutes of Health&#8217;s (NIH) guidelines in a specific pathogen-free facility at the Johns Hopkins University. All procedures were approved by the institutional animal care and use committee.
<p class="jov...

<ol>
	<li>Billingham, R. E., Brent, L., Medawar, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Actively+acquired+tolerance+of+foreign+cells.">Actively acquired tolerance of foreign cells.</a> Nature. 172 (4379), 603-606 (1953).</li><li>Murray, J. E. <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+(skin,+kidney,+heart)+and+the+plastic+surgeon.">Organ transplantation (skin, kidney, heart) and the plastic surgeon.</a> Plastic and Reconstructive Surgery. 47 (5), 425-431 (1971).</li><li>Liu, F., Kang, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+heart+transplantation+in+mice.">Heterotopic heart transplantation in mice.</a> Journal of Visualized Experiments. (6), e238 (2007).</li><li>Ruzza, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterotopic+heart+transplantation+in+rats:+improved+anesthetic+and+surgical+technique.">Heterotopic heart transplantation in rats: improved anesthetic and surgical technique.</a> Transplant Proceedings. 42 (9), 3828-3832 (2010).</li><li>Oldani, G., Lacotte, S., Morel, P., Mentha, G., Toso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+liver+transplantation+in+rats.">Orthotopic liver transplantation in rats.</a> Journal of Visualized Experiments. (65), e4143 (2012).</li><li>Lang, K. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunoprivileged+status+of+the+liver+is+controlled+by+Toll-like+receptor+3+signaling.">Immunoprivileged status of the liver is controlled by Toll-like receptor 3 signaling.</a> Journal of Clinical Investigation. 116 (9), 2456-2463 (2006).</li><li>Sun, Z., 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=Recruitment+of+host+progenitor+cells+in+rat+liver+transplants.">Recruitment of host progenitor cells in rat liver transplants.</a> Hepatology. 49 (2), 587-597 (2009).</li><li>Orlando, G., Soker, S., Wood, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Operational+tolerance+after+liver+transplantation.">Operational tolerance after liver transplantation.</a> Journal of Hepatolgy. 50 (6), 1247-1257 (2009).</li><li>Lodhi, S. A., Lamb, K. E., Meier-Kriesche, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Solid+organ+allograft+survival+improvement+in+the+United+States:+the+long-term+does+not+mirror+the+dramatic+short-term+success.">Solid organ allograft survival improvement in the United States: the long-term does not mirror the dramatic short-term success.</a> American Journal of Transplantation. 11 (6), 1226-1235 (2011).</li><li>Engels, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectrum+of+cancer+risk+among+US+solid+organ+transplant+recipients.">Spectrum of cancer risk among US solid organ transplant recipients.</a> Journal of the American Medical Association. 306 (17), 1891-1901 (2011).</li><li>Kasiske, B. L., Snyder, J. J., Gilbertson, D., Matas, A. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nephrotoxicity+of+immunosuppressive+drugs:+long-term+consequences+and+challenges+for+the+future.">Nephrotoxicity of immunosuppressive drugs: long-term consequences and challenges for the future.</a> American Journal of Kidney Diseases. 35 (2), 333-346 (2000).</li><li>Hu, X., 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=Chimeric+Allografts+Induced+by+Short-Term+Treatment+With+Stem+Cell-Mobilizing+Agents+Result+in+Long-Term+Kidney+Transplant+Survival+Without+Immunosuppression:+A+Study+in+Rats.">Chimeric Allografts Induced by Short-Term Treatment With Stem Cell-Mobilizing Agents Result in Long-Term Kidney Transplant Survival Without Immunosuppression: A Study in Rats.</a> American Journal of Transplantation. 16 (7), 2055-2065 (2016).</li><li>Shrestha, B., Haylor, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+rat+models+of+chronic+allograft+nephropathy:+a+review.">Experimental rat models of chronic allograft nephropathy: a review.</a> International Journal of Nephrology and Renovascular Disease. 7, 315-322 (2014).</li><li>Fu, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+transplantation+of+kidney+allografts+in+sensitized+rats+after+syngeneic+hematopoietic+stem+cell+transplantation+and+fludarabine.">Successful transplantation of kidney allografts in sensitized rats after syngeneic hematopoietic stem cell transplantation and fludarabine.</a> American Journal of Transplantation. 14 (10), 2375-2383 (2014).</li><li>Grau, V., 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=Immune+Complex-Type+Deposits+in+the+Fischer-344+to+Lewis+Rat+Model+of+Renal+Transplantation+and+a+Subset+of+Human+Transplant+Glomerulopathy.">Immune Complex-Type Deposits in the Fischer-344 to Lewis Rat Model of Renal Transplantation and a Subset of Human Transplant Glomerulopathy.</a> Transplantation. 100 (5), 1004-1014 (2016).</li><li>Vogelbacher, 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=Bortezomib+and+sirolimus+inhibit+the+chronic+active+antibody-mediated+rejection+in+experimental+renal+transplantation+in+the+rat.">Bortezomib and sirolimus inhibit the chronic active antibody-mediated rejection in experimental renal transplantation in the rat.</a> Nephrology Dialysis Transplantation. 25 (11), 3764-3773 (2010).</li><li>Fisher, B., Lee, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microvascular+surgical+techniques+in+research,+with+special+reference+to+renal+transplantation+in+the+rat.">Microvascular surgical techniques in research, with special reference to renal transplantation in the rat.</a> Surgery. 58 (5), 904-914 (1965).</li><li>Mahabir, R. 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The authors have nothing to disclose.

In this manuscript, we describe the surgical method for orthotopic KT in rats in detail, including all the necessary equipment needed to perform this procedure (Figure 5). In 1965, Fisher and Lee published the first report on KT in rats, which became the start of an exciting investigative field18. Since then, many modifications have been introduced to improve the reproducibility of this model. It has served as an effective animal model for studying ischemia-reperfusio...

Historically, the first transplant rejection studies were performed by Brent and Medawar using skin transplants in rodents1. It soon became clear that skin has distinct immunological features, making it a highly immunogenic organ that is different in rejection from other vascularized solid organs2. Rat studies of solid organ transplant rejection are habitually limited to heart, liver, and kidney transplants. Although each of these organs is suitable to study rejection, there are advantages and disadvantages to each of them. Heart transplants are often transplanted into the abdomen and anastomosed to the aorta and vena cava, with the recipient&#8217;s native heart in place3. This does not recreate human clinical, anatomical, and physiological conditions. Additionally, hearts are very sensitive to cold ischemia and have to be reperfused preferentially within 1 h in order to be able to recover their function4. Liver transplants are generally considered to be surgically more challenging and time-sensitive to perform. After removing the native liver, the donor liver has to be implanted and reperfused within 30 min as the recipients cannot last longer without a functioning liver5. The hepatic artery, portal vein, and especially the bile duct reconstruction requires refined surgical skills. Besides the surgical challenges, the liver is known to possess tolerogenic properties and rodents and humans can become operationally tolerant6,7,8. The kidney, unlike the aforementioned organs, can be transplanted in an orthotopic fashion, is known to be an immunogenic organ with consistent, reproducible rejection episodes (if not immunosuppressed), and allows for prolonged cold ischemia times of several hours. This makes the rat kidney transplant an ideal model for studying allograft rejection and tolerance.
Kidney transplantation (KT) is the preferred choice of treatment for patients with end-stage renal disease. Over the last few decades, short-term survival outcomes after KT have improved dramatically, but long-term survival outcomes are stagnant9. Conventional immunosuppressive regimens remain the standard anti-rejection therapy. However, the chronic use of immunosuppressive therapies causes significant morbidity and mortality, such as nephrotoxicity, diabetes, and secondary malignancies10,11,12. In the long-term, chronic antibody- and cellular-mediated rejection threaten graft survival, with limited therapeutic options available.
A major goal in transplantation is the induction of transplant tolerance in order to obviate the need for chronic immunosuppression. The rat KT model is a robust tool to investigate the immunological rejection process and to evaluate new approaches to immunomodulation and transplant tolerance. The rat also serves as a suitable model to study acute and chronic cell- and antibody-mediated rejection13,14,15,16,17. This surgical model has proven to be a reliable, reproducible, and cost-effective tool to study various aspects of allograft rejection and tolerance. It is often used to test novel tolerance-inducing protocols prior to undertaking expensive and cumbersome large-animal studies. Performing KT in rats requires extensive surgical training and expertise to reach survival rates of &#62;90%. In this manuscript and in the accompanying instructional video, we provide a step-by-step outline for orthotopic KT in the rat, as successfully performed for many years at our institution.
Prior to starting any procedure, donor and recipient selection is critical and depends on the nature of the experiment. Ideally, donors and recipients should weigh between 220&#8211;260 g and be between 8&#8211;12 weeks of age. Animals under 220 g have small-diameter arteries, veins, and ureters, making the anastomosis in the recipient particularly challenging. Minor blood loss can cause hypovolemia and lead to death in smaller animals. Animals heavier than 260 g display more fat around their vessels, and vessel isolation will require more operative time and increase the cold ischemia time.

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	<colgroup>
		<col />
		<col span="2" />
		<col />
	</colgroup>
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Buprenorphine HCL</td>
			<td style="width:167px;">Reckitt Benckiser Healthcare UK</td>
			<td style="width:167px;">NDC12496-0757-5</td>
			<td style="width:463px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dissecting forceps, curved</td>
			<td style="width:167px;">Zhenbang, China</td>
			<td style="width:167px;"></td>
			<td style="width:463px;">11cm Flat handle&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heparin sodium injection USP</td>
			<td style="width:167px;">Sagent Pharmaceuticals&#160;</td>
			<td style="width:167px;">NDC25021-400-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro-forceps, straight, smooth</td>
			<td style="width:167px;">Jingzhong, China</td>
			<td style="width:167px;">WA3010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro needle holder</td>
			<td style="width:167px;">Jingzhong, China</td>
			<td style="width:167px;">WA2010</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro vessel clamps</td>
			<td style="width:167px;">Jingzhong, China</td>
			<td style="width:167px;">WA40120</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro spring sciccor 1</td>
			<td style="width:167px;">ROBOZ</td>
			<td style="width:167px;">RS-5620</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro spring sciccor 2</td>
			<td style="width:167px;">F.S.T.</td>
			<td style="width:167px;">91501-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Micro spring sciccor 3</td>
			<td style="width:167px;">Zhenbang, China</td>
			<td style="width:167px;"></td>
			<td style="width:463px;">8.5cm Vannas,curved</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Prograf (Tacrolimus/FK506)</td>
			<td style="width:167px;">Astellas</td>
			<td style="width:167px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Rats</td>
			<td style="width:167px;">Charles River &#38; Taconic Biosciences&#160;</td>
			<td style="width:167px;">LEW/Crl &#38; DA-M&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Shaver</td>
			<td style="width:167px;">Wahl</td>
			<td>79600-2101</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Suture 4-0</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:167px;">J304H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Suture, 4-0&#160;</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:167px;">683G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Suture, 10-0&#160;</td>
			<td style="width:167px;">Ethicon</td>
			<td style="width:167px;">2820G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Syringes &#38; Needles</td>
			<td style="width:167px;">BD</td>
			<td style="width:167px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Thread, 8-0</td>
			<td style="width:167px;">Ashaway</td>
			<td style="width:167px;">75290</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ureteral cuff</td>
			<td style="width:167px;">Microlumen</td>
			<td style="width:167px;">160-1</td>
			<td>Polymide Tubing, Diameter 0.41 mm&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Venous cuff</td>
			<td style="width:167px;">Intramedic BD</td>
			<td style="width:167px;">7441</td>
			<td>PE-200 Non-radiopaque polyethylene tubing ID: 1.4 mm, OD: 1.9 mm</td>
		</tr>
	</tbody>
</table>

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.

We performed syngeneic (N = 5) and allogeneic kidney transplants (N = 5). Animals with a syngeneic transplant achieved long-term survival without any immunosuppressive treatment. Animals that received an allogeneic transplant without immunosuppression rejected their graft and succumbed to renal failure with a median survival of 8 days (Figure 4A). Mean serum creatinine increased modestly in the syngeneic group while it increased by 14-fold in the allogeneic...

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Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach

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.

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

Our laboratory at Johns Hopkins has been performing small animal kidney and liver transplantation with over two decades of experience. This specific protocol outlines critical steps that are necessary for successfully performing orthotopic kidney transplantation in rats. To increase the probability and reproducibility of success, our protocol includes three distinctive steps, perfusion of the donor kidney through the portal vein and the use of a cuff system to anastomose the renal veins and ureters.Demonstrating this procedure will be Dr.Le Qi, a graduate student in our laboratory. To begin this procedure, anesthetize the donor rat as described in the text protocol. Place the rat in a supine position and immobilize the limbs with sterile masking tape.Apply an eye lubricant and sterilize the surgical field as described in the text protocol. Prior to the first incision, make sure that the rat is adequately anesthetized by checking the absence of the toe pinch withdrawal reflex. Using scissors, start off by making a large longitudinal midline skin and muscle incision from the symphysis pubis to the xiphoid and enter the peritoneal cavity.Cover the intestine with a moist sterile gauze and shift it to the right lateral side of the abdomen exposing the aorta, vena cava, and left kidney. Apply one milliliter of preheated saline with a one cc syringe to keep the intestines and the abdominal organs moist and at a normal temperature. Apply a second moist gauze to cover and mobilize the stomach and spleen cranial to the kidney.Then add a small moist gauze to cover the exposed kidney. Use microsurgical dissecting forceps to isolate and mobilize the left renal artery and vein from the connective tissue and each other. Now isolate the left renal vein by cauterizing the left gonadal vein and isolate the left renal artery by cauterizing the adrenal artery.After that, mobilize the aorta and vena cava superior and inferior to the left renal pedicle by dissecting the connective tissue with dissecting forceps. Divide and mobilize the ureter from the connective tissue using dissecting forceps, then use micro scissors to make a diagonal incision at a length of approximately two centimeters measured from the renal pelvis. Insert a polyamide cuff halfway into the ureter and secure the cuff by placing a knot with 8-0 silk suture.Mobilize the left kidney by separating it from the perinephric fat using dissecting forceps or micro scissors. Leave the adipose capsule of the kidney attached and use that site for handling the kidney. Now administer 200 units of heparin using a syringe with a 27 gauge needle through the penile vein.Pressure the site of injection with a cotton swab for at least one minute to prevent bleeding. Identify the portal vein and inferior vena cava. Expose the inferior vena cava.Prepare to flush the kidney by injecting 50 milliliters of cold saline mixed with 500 units of heparin into the portal vein using a 16 gauge needle. Before flushing, cut the inferior vena cava at the infrahepatic level and cut the portal vein caudal at the needle insertion site to allow the blood to exit the circulation. Start flushing the kidney by gradually infusing the saline solution.Observe a color change of the kidney from dark red to a uniform gray or pale color. After flushing, ligate the renal artery and vein proximal to the aorta and vena cava. Place the flushed kidney in a Petri dish containing cold saline on ice before removing the donor's body from the surgical field.Once the kidney is in cold saline, fix and immobilize the handle of the vein cuff and gently pull the renal vein through the cuff. Then fix the renal vein over the cuff by placing three knots using 8-0 silk suture. To begin the recipient procedure, repeat the steps from the donor procedure up to heparin administration.Then place two atraumatic microvessel clamps on the left renal artery and vein proximal to the aorta and vena cava. Ligate the recipient renal vein proximal to the inlet of the kidney. Flush the renal vein with heparinized saline to remove all the remaining blood out of the vessel.Slide the ligated renal vein over the cuffed renal vein previously positioned in the donor kidney. Then secure the vein with an 8-0 silk suture. Maintain the same positional orientation when securing the renal vein over the cuff.Ligate the renal artery proximal to the inlet of the recipient kidney. Flush it with heparinized saline to remove any excess blood in the vessel. Now ligate the ureter at the level of the lower pole of the left kidney.Mobilize the kidney from the perinephric fat and remove the native kidney. Perform an end-to-end anastomosis of the renal artery with 8-10 interrupted sutures using a 10-0 nylon suture. Maneuver the artery by using the adventitial layer.Remove the vessel clamps to re-initiate the re-perfusion of the kidney. Start by removing the clamp on the vein followed by the clamp on the artery. Use a sterile cotton swab to add light pressure to any oozing areas around the anastomosis region.A few minutes should be sufficient to achieve a patent anastomosis. Briefly observe the kidney to assess for adequate perfusion. Immediately after re-perfusion, the kidney should change color and gradually regain its natural dark red color after a few minutes.Visible peristalsis of the ureter and onsite urine production are sometimes observed. Finish by inserting the exposed tip of the ureteral cuff into the recipient ureter and secure the recipient ureter with an 8-0 silk tie. In order to keep the donor and recipient ureters in position, tie off the ends of each ureter side to each other.Optionally, nephrectomize the right kidney by tying off the renal pedicle with a 3-0 silk suture. Remove all gauzes from the abdominal cavity and return all the organs to their natural position. Then squirt one milliliter of saline over the intestines to keep them moist.Close the abdomen by using a 4-0 absorbable suture on the rectus muscle and a 4-0 silk suture to close the skin layer in an interrupted fashion. Place the animal in a clean cage with access to ad libitum water and food and allow for recovery on a 37 degree Celsius heating pad. Observe the recovery for one to two hours before returning the animal back to the animal facility.Animals with a syngeneic kidney transplant achieved long-term survival without any immunosuppressive treatment. Conversely, animals that received allogeneic kidney transplant without immunosuppression rejected their graft and succumbed to renal failure with a median survival of eight days. Mean serum creatinine increased modestly in the syngeneic group while it increased by 14-fold in the allogeneic group.Upon explantation, the macroscopic view of the syngeneic kidney allograph did not show any abnormalities. The kidney color and internal structures remained intact. In contrast, kidney allographs of rejected animals presented red hemorrhagic patches with the destruction of the internal structures.Hematoxylin and Eosin stains of syngeneic grafts shown thin glomerular capillary loops with normal numbers of endothelial and mesangial cells. Rejected allographs displayed destroyed glomerular structures with signs of inflammation and tubulitis. CD8 positive staining was performed to confirm T-cell mediated rejection.While syngeneic allographs showed very few CD8 positive T-cells, the rejected allographs showed a significantly higher number of CD8 positive cells in and around the glomeruli and tubuli confirming T-cell mediated rejection. This model provides significant insight into the immunological phenomena of rejection, tolerance, and donor-host interactions paving the way for the development of therapeutical clinical interventions in solid organ transplantation.

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15.2K Views.  Johns Hopkins University School of Medicine. Our laboratory at Johns Hopkins has been performing small animal kidney and liver transplantation with over two decades of experience. This specific protocol outlines critical steps that are necessary for successfully performing orthotopic kidney transplantation in rats. To increase the probability and reproducibility of success, our protocol includes three distinctive steps, perfusion of the donor kidney through the portal vein and the use of a cuff system to anastomose the renal veins and u...