Supplementary Figure 1: Experimental design and ex-vivo rat kidney perfusion techniques b

(1)

Cold storage (18 or 4 hours)

Supplementary Figure 1: Experimental design and ex-vivo rat kidney perfusion techniques

b DCD Kidney grafts during different machine perfusion approaches Normothermic

oxygenated perfusion Hypothermic oxygenated Perfusion (HOPE)

Kidney flush (saline + 1U Heparin/ ml, 20 °), canulation of ureter Laparotomy, cannulation of ureter

CS + cuff Cuff placement during minimal cold storage (10 Min) Kidney flush and Stent placement for Perfusion

Endpoints d

a

CS+cuff Kidney Trans-

plant

HOPE 1hr

Kidney Trans-

plant

CS+cuff

Study groups 1a) No Injury (Baseline)

2) DCD + CS

3) DCD + CS + HOPE

4. DCD + CS + HNPE 5. DCD + CS + Normotherm

HNPE 1hr

plant

CS+cuff

Normo-1hr therm

plant

CS+cuff

plant

CS+cuff

Initial Preservation Treatment Transplantation Injury

Hypothermic nitrogena- ted Perfusion (HNPE)

c

plant

CS+cuff

1b) No Injury + normotherm or

+ hypotherm

Perfusion Parameter Normotherm HOPE HNPE

PO2 (kPa) 55-65 80-100 <2

Flow (ml/min) 2.057 ± 0.299 0.75 ± 0.22 ml/min 0.82 ± 0.13

Pressure (mmHg) < 75 < 18 < 18

Temperature (°C) 37 4 4

Perfusate

Isolated, diluted erythrocytes, Ringers Solution (HCT: 20%),

additives

⁽⁶⁾

KPS-1 KPS-1

IRR (intrarenal resistance,

mmHg/ml/min/100g) 0.30

Perfusion Parameter

Plasma + Tissue analysis

0hrs 1 day 1 week Kidney

Transplantation At the end of

Preservation/Machine perfusion 1h nor-

mo- or hypo- therm

Laparotomy, opening of diahpragma, awaiting cardiac arrest and 30 minutes asystolic warm ischemia, (no heparin)

DCD: a period of 30 minutes warm in situ asystolic ischemia

(2)

Supplementary Figure 1: Experimental design and ex-vivo rat kidney perfusion techniques b