Murine Kidney Transplant Technique

The overall goal of this procedure is to demonstrate a modified arterial anastomosis technique for kidney transplants in the mouse. This is accomplished by first carefully dissecting the relevant vessels in the donor. The second step is to create a heel and toe cuff of the donor abdominal aorta adjacent to the renal artery.

Next, this heel and toe cuff is used to form the arterial anastomosis in the recipient. Ultimately, this technique should result in very low instances of arterial thrombosis. This video will show you how to create an arterial heel and toe cuff to facilitate kidney transplants in the mouth.

The main advantage of this technique of resisting methods is that the blood flow path from the recipient aorta into the donor renal artery is straightened, resulting in a reduction of platelet activation and thrombosis formation Using sterile technique under an operating microscope, harvest the donor's kidney, perform properly. The harvest time will be 15 to 20 minutes. Remove the fur and then restrain the animal.

Prepare the surgical site by washing the skin with povidone iodine, drape the mouse and begin the surgery with a two centimeter midline abdominal incision. Retract the bowel onto the chest and wrap it in moist gauze. Keep the gauze moist throughout the procedure.

Now, find the great abdominal vessels and mobilize them. Any lumbar branches should be either cauterized or ligated with 10 zero nylon suture. Next bluntly, dissect a two millimeter division of the inferior vena cava and the aorta distal to the left renal vessels.

Then ligated and divide any small arterial and venous branches that attach to the renal vessel. Continue the blunt dissection to separate the renal vein from the renal artery, thus allowing for the later creation of a Carroll patch at the proximal end of the renal vein, this patch will be used for the formation of the venous anastomosis. Next like it and divide the left adrenal vessels to access the supra renal aorta.

Once accessed, loop six zero silk. Suture around the supra renal aorta for later use. Now mobilize the kidney vessels and ureter from the surrounding fascia.

Rotate the kidney to the right on the posterior side, ligate and divide any vascular branches focusing in on the ureter. Carefully free the ureter from the surrounding fascia. Do not disturb the renal hilum and preserve the eretic vessels.

Next, divide the ureter at the level of the ductus deens. The kidney can now be recovered. First, slowly inject 300 units of heparin into the distal IVC to heparinized the donor.

Second, tie down the six zero suture around the supra renal aa. Then perfuse the left kidney from the abdominal aorta, injecting 0.8 milliliters of heparin saline solution at 100 units per milliliter. After the perfusion, immediately create the venous caryl patch, thereby avoiding backflow into the kidney.

Retract the renal vein toward the kidney to reveal the AA and renal artery. Now divide the aorta adjacent to the renal artery. Make a heel and to cuff.

Then if zero minutes of cold ischemia is required, transfer the recovered kidney into a prepared recipient. Alternatively, transfer the organs to a four degree Celsius storage chamber and then euthanize the donors as indicated in the text protocol. This procedure will take 35 to 45 minutes as done for the donor anesthetize.

The recipient also supplement the initial bolus as needed. Provide the recipient with ophthalmic cream. Then give the animal an injection of buprenorphine and continue its administration after the procedure, after making the initial incision and displacing the bowel as done with the donor, proceed with a right nephrectomy, ligate the renal artery and vein with six zero suture ligate and divide the ureter proximal to the kidney with six zero suture.

Then excise the kidney distal to the vascular suture. Now below the renal vessels, isolate the AA and IVC then superior to the anastomosis site. Place a four zero cotton tie around the aorta and IVC do the same inferior to the site.

Any lumbar vessels found in the region should either be cauterized or ligated with 10 zero nylon suture tightly not the suture that is inferior and then tightly not the suture that is superior. This will retain some blood in the aorta facilitating the AOR autotomy. Perform the atomy with a 30 gauge needle, then extend the incision to be about two millimeters long using fine micro scissors.

The next step is to make an endo anastomosis from the donor heel and toe cuff to the recipient's aorta. First, put a stay stitch in the heel of the donor aorta with 10 zero suture. Continue this through the inferior end of the incision in the recipient's aorta and tie off.

Second, place a 10 zero suture in the superior end of the aortic incision and continue this through the toe of the donor cuff and tie off. Thirdly, starting at the superior end, make a runny suture in the lateral wall of the aorta and tie off against the previously placed inferior stay stitch. Then flip the kidney and continue a running suture along the medial wall and tie off against the superior stay stitch.

Complete the venous anastomosis by puncturing the IVC using the needle and then extending the incision with fine micro scissors. Using 10 zero nylon, tie the donor renal vein to the inferior corner of the IVC. Place another 10 zero suture at the superior end of the IVC and donor vein and make a running suture along both sides.

As with the arterial anastomosis, aversion of the tissue edges helps with seal and healing. Now, check your work. Opposing walls should not be caught by the stitches or blood flow will be too constricted.

Another vitally important factors ensuring that the tension of the anastomotic suture line is optimal too loose and there will be irreversible leaking too tight and there will be stricture to flow. If on the arterial side, this will result in poor profusion of the graft. If on the venous side a congested kidney will result.

Once the anastomosis are complete, release the distal cotton tie to establish venous flow. Then gradually loosen the proximal tie and wash for hemostasis at the arterial anastomosis. When both anastomosis are confirmed as secure, here's the recipient's bladder with a 20 gauge needle making two holes pass forceps through the holes and pull the donor ureter through the bladder.

Then anchor the proximal end of the ureter to the bladder wall with 2 10 0 sutures. Trim the excess length of protruding ureter so it retracts within the bladder. Then close the other hole in the bladder with 10 zero suture.

Complete the surgery by replacing the bowel, closing the abdomen in two layers with running five zero silk suture and injecting one microliter of saline into the abdomen before closing. After closing the abdomen inject another 0.8 milliliters of saline subcutaneously and recover the animal on a warm blanket In one year of data collection, this technique significantly reduced the incidence of arterial thrombosis from 35%to 0%and this rate of 0%thrombosis has been maintained since immunohistochemistry was performed on a non-transplant kidney and a kidney transplanted. Using the described technique, a periodic acid shift stain revealed normal proximal tubular cells in the control and donor.

The cells are cuboidal with a clear cytoplasm and a round light nucleus in the middle of the cell for both the arrows. Point out evidence of a mild brush border injury, but the majority of brush borderers are regular and well preserved. After watching this video, you should have a good understanding of how to perform an arterial heel and toe cup anastomosis that will result in reduced instances of thrombosis and more successful kidney transplants.