Chapter 5 138 MATERIALS & METHODS Ethical approval Ethical approval (number B19.019) for experiments with discarded donor kidneys was granted by the Medical Ethical Committee of the Leiden University Medical Center (LUMC) and University Medical Center Groningen (UMCG). Research consent was obtained from the relatives of all donors prior to organ retrieval. Procurement, preparation of the kidney, and sample drawing The included kidney grafts (N=8) were procured from deceased donors according to Dutch national guidelines and were deemed untransplantable because of procurement-related factors or factors determined prior to retrieval (specified in Table 1). After in situ flushing of the abdominal organs with cold University of Wisconsin (UW) preservation solution, the kidneys were retrieved, preserved by either static cold storage (SCS) or HMP, and transported to the participating centers (LUMC or UMCG) where NMP was initiated and performed up to 6 hours (Figure 5). Upon arrival in the participating centers, kidney grafts were inspected and prepared for connection to the NMP circuit under sterile conditions, while the perfusion machine (Kidney Assist, Organ Assist, Groningen, The Netherlands) was primed. The NMP setup was primed as previously described in the PROPER study 24. Kidneys were subjected to subsequent NMP when the Cold Ischemia Time (CIT) did not exceed 24 hours at arrival. Perfusate samples were drawn prior to (0) and after 60 / 180 / 360 minutes of NMP, centrifuged at 3700 rpm at room temperature, and the supernatant was stored at -80 0C. Figure 5 – Schematic overview representing procurement, transportation, preparation of the kidneys, and sample drawing.
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