Modeling Preservation Injury in Porcine Donor Kidney: A Procedure to Induce Graft Dysfunction by Subjecting Kidney to Prolonged Cold Ischemia

Preservation of donor graft organ deprived of its blood supply is crucial for maintaining its viability ex vivo until transplantation. Cold preservation of organ for extended periods impairs organ function, causing preservation injury and impacting transplantation outcomes. 

To induce organ preservation injury, begin with a freshly excised porcine donor kidney with its renal artery, renal vein, and ureter segment. Insert and secure a catheter to the renal artery. Connect the catheter to a chilled organ preservation solution-containing reservoir. Perfuse the kidney with the organ preservation solution to remove blood remnants from the organ through the venous end. Remove the catheter.

Transfer the kidney to a sterile organ bag containing chilled organ preservation solution which provides nutrients for reduced energy demands of the organ. Store the organ maintaining low temperatures for a prolonged duration, using a computer-controlled cooling system. The viscosity of the preservation solution facilitates a shorter cooling time for the organ to reach a low temperature.

The lack of blood flow to the organ, ischemia, interrupts oxygen and nutrient delivery, affecting cellular energy metabolism. Prolonged cold ischemic time causes severe cellular energy depletion and ultrastructural changes, leading to cell death. This kidney preservation injury model can be used to study clinical organ transplantation challenges and improve organ preservation techniques. 

Immediately after retrieval, use a standard 14-gauge peripheral catheter to cannulate the renal artery, and flush the kidney with at least 500 milliliters of ice-cold organ preservation solution. After the flush, wrap the graft in sterile organ bags for storage in organ preservation solution, with a target cold ischemic time of 24 hours at 4 degrees Celsius using a computer-controlled cooling circuit.