Technical Refinement of a Bilateral Renal Ischemia-Reperfusion Mouse Model for Acute Kidney Injury Research

Acute kidney injury is a major risk factor associated with high mortality in patients with cardiac arrest after return of circulation. This procedure can be performed to induce acute kidney injury. This protocol is to induce renal ischemia reperfusion accurately, securely, and consistently.

Demonstrating the procedure will be Huang Cheng-Wei, an assistant from my laboratory. To begin, place an anesthetized eight-week-old C57BL/6 male mouse in the prone position on a surgery platform with an electric blanket to maintain body temperature. Next, touch the back, find the lumbar spine, and look for costovertebral angles below both sides of the last rib.

Apply hair removal lotion to shave and scrub the hair near both sides of the costovertebral angle region. Disinfect the surgical area using a Betadine solution and 75%alcohol using cotton balls. Use fine tip forceps to gently lift the skin below the left costovertebral angle.

Then, using scissors, create a one-centimeter oblique dorsolateral incision along the skin tension lines from the lumbar midline on the left flank. Next, transect the muscle wall of the left flank to visualize the left kidney. Push and separate the left kidney carefully from the surrounding tissue with forceps.

Then, identify the renal pedicle. Clamp the left renal pedicle with a microvascular clip. Confirm the ischemia by a visible change in kidney color from pink to dark red.

To avoid desiccation during clamping, cover the kidney with sterile saline-wet cotton balls. After 25 minutes, open the left microvascular clip to start reperfusion of the left kidney. Return the kidney to the abdominal cavity.

Close the abdominal cavity with 6/0 absorbable suture materials. Observe the mouse until it begins to move freely and feed. Renal ischemia after renal pedicle clamping is shown here.

A kidney color change from pink to dark red reveals that the kidney perfusion has become inadequate. Compared to the controls, serum levels of blood urea nitrogen and creatinine increased two days after renal reperfusion. This technique paves a way for researchers to explore novel therapeutic approaches against acute renal injury.