Preparation of Decellularized Rat Kidney: A Procedure to Generate Acellular Vascularized Whole-organ Scaffold from a Harvested Rat Kidney
Transcripción
Harvest the left kidney with the abdominal aorta and inferior vena cava. Ligate the ureter, thoracic aorta, superior vena cava, and branches of the abdominal aorta. Keep the organ hydrated in DPBS in a 10-centimeter Petri dish. Cannulate the abdominal aorta and inferior vena cava with a 23-gauge catheter.
To remove residual blood, connect the cannula with a peristaltic pump and wash the organ with 500 milliliters of DPBS and 16 units per milliliter heparin for 90 minutes at 5 rpm and 37 degrees Celsius. To decellularize the kidney, perfuse it with 1% Triton X-100 for 3 hours and then with 0.75% SDS solution for 6 hours at a constant pressure of 40 milliliters of mercury.
To remove residual SDS, perfuse the sample with 1% penicillin in distilled water for 18 hours and then with sterile DPBS and 16 units per milliliter heparin for 90 minutes.
