Murine Renal Transplantation Procedure


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Renal transplantation in mice is a technically challenging procedure that requires careful post-operative care and treatment for success.

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Wang, J., Hockenheimer, S., Bickerstaff, A. A., Hadley, G. A. Murine Renal Transplantation Procedure. J. Vis. Exp. (29), e1150, doi:10.3791/1150 (2009).


Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell et al over 30 years ago (1) and refined by Zhang et al years later (2), few people in the world have mastered this procedure. In our laboratory we have successfully performed 1200 orthotopic kidney transplantations with > 90% survival rate. The key points for success include stringent control of reperfusion injury, bleeding and thrombosis, both during the procedure and post-transplantation, and use of 10-0 instead of 11-0 suture for anastomoses.

Post-operative care and treatment of the recipient is extremely important to transplant success and evaluation. All renal graft recipients receive antibiotics in the form of an injection of penicillin immediately post-transplant and sulfatrim in the drinking water continually. Overall animal health is evaluated daily and whole blood creatinine analyses are performed routinely with a portable I-STAT machine to assess graft function.


Donor Organ Harvest (2):

  1. Anesthetize the donor using 65mg/kg pentobarbital IP; anesthesia is maintained by Isoflurane (2-3%).
  2. Make a midline incision from sternum to pubis to enter the donor abdomen and expose the left kidney by moving the intestine laterally to the right side.
  3. Isolate the left kidney by ligating and dividing the adrenal and testicular vessels with an 8-0 silk suture.
  4. Mobilize the aorta above its junction with the renal artery and vein.
  5. Mobilize the inferior vena cava (IVC) above and below its junction with the renal artery and vein.
  6. Ligate the aorta below the renal artery and vein with an 8-0 silk suture to minimize bleeding.
  7. Dissect the left ureter from the renal hilum to the bladder.
  8. Expose the ureterovesical junction using caudad retraction on the bladder dome.
  9. Excise a small, elliptical patch of bladder containing the left ureterovesical junction, used for urinary reconstruction via a bladder-to-bladder anastomosis. The right ureter is not included with the bladder.
  10. Ligate the IVC below the renal artery and vein with an 8-0 silk suture.
  11. Ligate the aorta above the renal artery and vein, and slowly perfuse the graft in situ with 0.2-0.4ml cold, heparinized Ringer's lactate solution via the infrarenal aorta.
  12. Ligate the IVC above the renal artery and vein with an 8-0 silk suture.
  13. Transect the renal vein at its junction with the IVC.
  14. Divide the aorta obliquely, approximately 2mm below the renal artery.
  15. Remove the kidney and its vascular supply with ureter attached to the bladder patch en bloc. Store in Ringer's lactate solution on ice.
  16. Euthanize the donor mouse by cervical dislocation under anesthesia.

Recipient Transplantation (2):

  1. Anesthetize the recipient with 65 mg/kg pentobarbital IP and maintain the anesthesia by Isoflurane (2-3%).
  2. Make a midline incision, cover the intestine with wet gauze and carefully retract it to the left side.
  3. Ligate the right ureter, renal artery and vein, and remove the right kidney.
  4. Carefully isolate and cross-clamp the infrarenal aorta and IVC with two 4-mm microvasular clamps after ligating the lumbar branches.
  5. Place a 10-0 nylon suture through a full thickness of aorta and retract to make an elliptical aortaotomy by a single cut (about one-fifth of the diameter of the vessel).
  6. Make a longitudinal venotomy by piercing the IVC with a 30 gauge needle and extend appropriately using microscissors.
  7. Irrigate both the aorta and IVC with heparinized saline to clear intraluminal blood or clots.
  8. Remove the donor kidney from the ice and place it intra-abdominally in the right flank of the mouse. The donor kidney is kept moist with a cold saline-soaked gauze pad.
  9. Place two stay sutures at the proximal and distal apex of the recipient’s aortaotomy with the donor’s aortic cuff.
  10. Make an end-to-side anastomosis between the donor aortic cuff and the anterior wall of the recipient’s aortaotomy, using 2 ~ 3 continuous 10-0 nylon sutures in the outside of the aorta.
  11. Turn the donor kidney graft over to the left flank of the recipient. repeat the previous procedure between the donor aortic cuff and the posterior wall of the recipient’s aortaotomy
  12. Perform an end-to-side anastomosis between the donor renal vein and the posterior wall of the recipient’s IVC, using 4 to 5 continuous 10-0 nylon sutures in the inside of the IVC. and then the anterior wall of IVC and donor’s renal vein are closed with 4 ~ 5 continuous sutures outside the IVC.
  13. The anastomosis of both vein and artery must be finished within 20 minutes.
  14. Rinse the graft with cold saline several times during anastomosis.
  15. Divide the dome of the recipient bladder, and stop bleeding with cautery.
  16. Make a small cut on the dome.
  17. Place two stay sutures of 10-0 nylon to anastomize the donor’s small bladder patch with the recipient’s bladder dome. Each side of anastomosis is sutured with 8 ~ 10 continuous stitches.
  18. Close the abdomen in two layers with continuous 4-0 sterile, synthetic absorbable vicryl suture.
  19. Give recipient mice 0.4ml warm saline during surgery by intermittent injection IV.

Post-operative Care of Recipient

  1. After closing the abdomen, a single intramuscular injection of penicillin (500U/10g) is administered (2).
  2. In addition, recipients receive 0.05 mg/kg buprenorphine intramuscularly on day 0 and day 1 post surgery for pain. Following surgery, transplant recipients receive water containing sulfatrim (100mg/kg) until the experiment is terminated.
  3. For the first 24h postoperatively mice are kept in a temperature controlled incubator.
  4. Mice usually recover from anesthesia within 1 hour of the operation and are given regular food and water ad libitum.
  5. Skin sutures are removed 2 weeks after the surgery.

Recipient Nephrectomy

  1. Anesthetize the recipient 4 ~ 7 days post kidney transplantation with 65 mg/kg pentobarbital IP; anesthesia is maintained by Isoflurane (2-3%).
  2. Open the abdomen via a midline incision.
  3. Cover the intestine with saline-soaked gauze and carefully retract it to the right side.
  4. Remove the left kidney after ligating the left ureter, kidney vein and artery.
  5. Close the abdomen in two layers with continuous 4-0 synthetic absorbable vicryl suture.

Whole Blood Creatinine Measurements

  1. Weekly to biweekly blood collection of less than 0.2 cc is performed either submandibularly without anesthesia or retro-orbitally under Isoflurane anesthesia.
  2. Quantitative whole blood creatinine levels are determined using an I-Stat Portable Clinical Analyzer (Heska Corp., Fort Collins, CO).
  3. Conventional units (mg/dl) are converted to SI units of micromol/L by multiplying the conventional units by 88.4.

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We have performed approximately 1200 cases of renal transplantation in mice with a technical success rate of >90%. Compared to heart transplantation, the success rate of kidney transplantation is usually low. However, our laboratory has maintained a high success rate (> 90%) of kidney transplantation. The key points for success are to reduce reperfusion injuries to both donor kidney and recipients, and minimize bleeding and thrombosis throughout the procedure and post-operatively. The following factors may be most important: (a) reduce bleeding in the recipient during surgery as much as possible through careful and gentle operation; (b) minimize warm ischemic and cross-clamping times during donor kidney preparation and recipient transplantation; (c) use a small donor bladder patch to prevent necrosis, insufficient blood supply and bladder leakage post-operation; (d) delay nephrectomy of the native kidney until 4 ~ 7 days post-transplantation to allow the transplanted kidney to recover function following surgery; (e) perform anastomoses (between the donor aortic cuff and the anterior wall and between the donor aortic cuff and the posterior wall of the recipient’s aorta) outside of the aorta to reduce the incidence of thrombosis; (f) keep recipient mice warm for at least for 24 hours following surgery; (g) give antibiotics to recipient mice post-operatively to prevent potential infections.

In this protocol, we use 10-0 instead of 11-0 suture without affecting survival rate. Although the 11-0 suture is ideal and used in many transplantation laboratories, we feel that 10-0 suture is sufficient for use in kidney transplantation as long as the factors described above are well controlled.

The removal of the remaining native kidney is essential so that decline in kidney graft function can be monitored by serum creatinine measurements. All kidney graft recipients undergo nephrectomy of the left native kidney between 4 and 7 days post kidney transplantation. Nephrectomy is not performed at the time of kidney transplantation because recipients characteristically suffer ill health or do not survive when both procedures are conducted at the same time. A 4 to 7 day lag between procedures permits the kidney graft to recover from ischemia and reperfusion injury before the native kidney is removed. Kidney graft function is assessed by daily examinations of overall animal health and blood creatinine levels. For reference, a non-transplanted, normal mouse has a creatinine level of about 20 micromol/L. Renal allograft rejection is suspected when the recipient mouse shows signs of illness and the creatinine level are elevated near 100 micromol/L at which time grafts are harvested for histopathological analyses.

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Experiments on animals were performed in accordance with the guidelines and regulations set forth by The Ohio State University IACUC.


These studies were supported in part by funding by NIH R01 AI36532 to GAH. We would like to acknowledge the outstanding support in transplant pathology provided by Dr. Tibor Nadasdy, and the invaluable guidance and foresight provided by the late Dr. Charles G. Orosz in initiating this research effort.


Name Type Company Catalog Number Comments
i-STAT Other HESKA Cat 5101
i-STAT Cartridge Other HESKA CAT 5110-CREA
Sulfatrim Other Actavis R02109
Isoflurane Other NOVAPLUS CA-0474
10-0 Suture Surgery Sharpoint AK0100



  1. Russell, P. S., Chase, C. M., Colvin, R. B., Plate, J. M. D. Kidney transplants in mice. An analysis of the immune status of mice bearing long-term, H-2 incompatible transplants. JExpMed. 147, 1449-1468 (1978).
  2. Zhang, Z., Schlachta, C., Duff, J., Stiller, C., Grant, D., Zhong, R. Improved techniques for kidney transplantation in mice. Microsurgery. 16, (2), 103-109 (1995).



  1. We are a studygroup from the Netherlands, performing murine experiments to determine the effect of a new therapy on the ischemia/reperfusion injury on kidneys. We have experience with mice models, but we have to compose our surgical instrument set. Is it possible to give the manufacturer and catalog number of the 4-mm microvasular clamps?
    Thanks in advance.

    Kinds regards

    Posted by: Anonymous
    August 12, 2009 - 9:48 AM
  2. Here is 4-mm micro vessel clamps information:

    Vendor: Biomedical Research Instruments.

    Catalog number: 14-1000.

    Posted by: Anonymous
    August 12, 2009 - 5:22 PM
  3. Good job and congrats on your procedure. May I know what type needle holder are you using? The curve is pretty god.


    Posted by: Reza N.
    September 19, 2009 - 12:56 AM
  4. Here is needle holder information:
    Vendor: Biomedical Research Instruments.
    Catalog number:²0-1050

    Posted by: Anonymous
    September 23, 2009 - 9:45 AM
  5. Hello, I have a question about the binocular magnification that you use in this procedure.
    Thanks in advance and good luck with your experiments.

    Posted by: Anonymous
    March 2, 2010 - 10:09 AM
  6. We use the eyepiece 1².5x.

    Posted by: Anonymous
    March 3, 2010 - 5:50 PM
  7. hallo,
    Is it possible to give the manufacturer and catalog number of the tweezers that you used and the catalog number of thesewing thread
    thank you very much and goodluck

    Posted by: Anonymous
    March 21, 2010 - 8:35 AM
  8. Here is tweezers information:
    Pattern #5 Vendor: ROBOXZ, Catalog number: RS-5045,
    Pattern #7 Vendor: ROBOXZ, Catalog number: RS-5047,
    10-0 Suture information:
    Vendor: Covidien,
    Catalognumber: N²51²

    Posted by: Anonymous
    March 23, 2010 - 2:40 PM
  9. We just start kidney transplant for genetic question. We have many problem coming. May I can visit your lab. to see you to transplant one time.

    Posted by: Anonymous
    July 13, 2011 - 10:48 AM
  10. Hi again Ben,

    Sorry sent that last reply before its time. Jiao-Jing's e-mail address is

    Posted by: Anonymous
    July 13, 2011 - 12:06 PM
  11. Hi Ben,

    Unfortunately, we no longer do mouse renal transplants in our lab. The person who did the transplants (Jiao-Jing Wang) is now at Northwestern University. I do not know whether you can observe here there, but you should contact her and ask. She has over 10 years of experience in mouse renal transplantion and is very skilled at this technique. Her e-mail address is

    is currently at

    Posted by: Anonymous
    July 13, 2011 - 12:02 PM
  12. Hi,

    Very nice video. Helps to better understand when learning this difficult technique.

    I have few questions please....

    1. What models have you used in these transplants?
    ². Did you use immunosuppression for allogenic transplants? If so, what drugs and how long?
    3. What is your survival rate?
    4. What were the common complications?

    Thank you,

    Posted by: Anonymous
    October 24, 2011 - 1:53 PM
  13. Hi Kumar,

    We have used this mainly for DBA/² renal allografts into C57BL/6 recipients. In these experiments, the recipients are primed with a DBA/² skin allograft, as DBA/² renal allografts into naive C57BL/6 recipients are spontaneously accepted. With sensitization, renal allografts in this model survive about 10 days, a time which can be extended indefinitely with a 3 day course of CD4 depletion. In contrast, depletion of CD8 cells has no effect on renal allograft survival. Another model in which we have extensive experience is the BALB/c into C57BL/6 model in which renal allografts are spontaneously rejected (MST ~100 days). Again, CD4 depletion promotes indefinite survival of renal allografts whereas CD8 depletion has no effect. Our mortality rate is less than 10% but you should expect it to approach 50% in the beginning. Our microsurgeon, Jiao-Jing Wang, has extensive experience with this technique, so is way ahead of the learning curve. A common problem even in the hands of Ms. Wang is hydronephrosis but the cause of most mortality is not diagnosed.


    Posted by: Anonymous
    October 24, 2011 - 2:31 PM
  14. Hi Gregg,

    Many thanks. Your information helps a lot.

    Your model of BALB/c to C57BL/6 seems to tolerate the kidney for more than three months without immunosuppression. That seems to be a good model. I am looking for a model which will stay alive for atleast 1 month without immunosuppression as it will be another factor which we wish to avoid in our study at present.

    I have done many transplants in rats and am starting it in mouse now.

    I read in an abstract that some strains accept all grafts with good acceptance but the article did not say which ones. So for now, I am planning to use C57BL/6 as both donor and recipients. We also have FVB and C3H but I did not come across any transplants between them.

    In rats, I had 40% hydronephrosis when I used to insert the ureter directly into the bladder but after switching to bladder patch technique as in the video, it is less than 5%.

    Thank you once again.

    Posted by: Anonymous
    October 24, 2011 - 4:21 PM
  15. use interrupt suture on ureter to bladder

    Posted by: zhonglin w.
    January 11, 2015 - 1:34 PM
  16. I am Gengwen from University of Wisconsin Madison. I started doing mice renal Tx 3 months ago and have done 40 operations. But unfortunately, I still didn't get a survivor. Recently, I can get the transplanted kidney perfused well and very little bleeding during the operation. But the animal still can not survive even ²4 hours. The  clamping time is about 30 minutes. The weight of mice is ²5-30 grams. The species is B6 or FVB.
     I saw your video on the Internet and I know you have over 10 year experience over this operation. So I wish you can help me analyze this problem and give me some advice.

     Thank you,


    Posted by: Anonymous
    February 24, 2012 - 1:43 PM
  17. Dear
    we are beginning the work of experimental renal transplantation in rats, I wonder what the most suitable material to start and what is your experience in the treatment of infections

    Posted by: Claudia C.
    August 4, 2014 - 12:06 PM
  18. You may inject penicillin after surgery or put antibiotics into water bottles for daily drink

    Posted by: Jiao-Jing W.
    August 8, 2014 - 10:12 AM

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