Overview
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.
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Materials
| Name | Company | Catalog Number | Comments |
| 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 | EH7694H | Skin |
