In this video, we describe a technique to induce preservation injury via the prolonged cold ischemic storage time in porcine donor kidney.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Back-table and organ preservation
After graft retrieval, immediately cannulate the renal artery using a standard 14 G (orange) peripheral catheter and fix it using a tourniquet prepared from 3-0 polyglactin.
Rinse the kidney with a cold organ preservation solution.
After flushing with 500 mL of organ preservation solution, remove the arterial cannula, wrap the kidney graft in sterile organ bags and store in organ preservation solution with a target cold ischemic time (CIT) of 24 h at 4 °C using a computer-controlled cooling circuit. NOTE: A brief post-preservation flush is recommended using 500 mL of 4 °C normal saline solution.
Açıklamalar
The authors have nothing to disclose.
Materials
Organ preservation solution (e.g. HTK)
Dr. Franz Köhler Chemie GmbH, Bensheim, Germany
Should be decided based on preference and experimental design
Organ preservation
Vessel loops
Medline International Germany GmbH,Kleve, Germany
VLMINB
Hold and adjust the vessel
Sterile organ bag
Raguse Gesellschaft für medizinische Produkte, Ascheberg, Germany
800059
Organ preservation
Micro steel needle holder straight 0,5mm, with spring lock
ASANUS Medizintechnik GmbH, Tuttlingen, Germany
MN1324D
Microsurgical needle holder
3M™ Standard Surgical Mask 1810F
3M Deutschland GmbH, Neuss, Germany
3M-ID 7000039767
Surgical mask
VICRYL – ligatures Sutupak purple braided, 3-0
Johnson & Johnson Medical GmbH – Ethicon Deutschland, Norderstedt, Germany
V1215E
Threads for ligature
Prolene 3-0,blue monofil,FS1 needle
Johnson & Johnson Medical GmbH – Ethicon Deutschland, Norderstedt, Germany
Modeling Preservation Injury in Porcine Donor Kidney: A Procedure to Induce Graft Dysfunction by Subjecting Kidney to Prolonged Cold Ischemia. J. Vis. Exp. (Pending Publication), e20913, doi: (2023).