Normothermic perfusion of pig kidney

First, the pig is sedated. After intubation and general anaesthesia, a midline laparotomy is performed and the kidney is freed from the surrounding tissue. Then the renal artery and vein are ligated and the kidney is removed. On the back-table, the kidney is placed on melting ice and the renal artery is canulated with an armoured vascular cannula. The ureter is also cannulated with a small tube. Then the kidney is flushed with a cold preservation solution via the renal artery at a hydrostatic pressure of 100 cm H2O to rinse out any blood. In the meantime the normothermic perfusion circuit has been primed with the perfusion solution (Table S1) and heated to 38°C. The kidney is mounted on the circuit by connecting the arterial cannula with the inflow cannula of the circuit. The tube in the ureter is connected to a collection bag allowing urinary drainage. Then the arterial inflow cannula is opened and the kidney is perfused. This is noticeable by pinking up of the kidney once reperfusion is established.首先对实验猪实施镇静处理。完成气管插管与全身麻醉后，行正中剖腹术，将肾脏与其周围组织游离。随后结扎肾动脉与肾静脉，摘除肾脏。在离体器官修整台上，将肾脏置于碎冰之上，采用加强型血管套管（armoured vascular cannula）对肾动脉进行插管，同时以细管对输尿管实施插管。随后以100 cm水柱的静水压经肾动脉向肾脏灌注冷保存液，以冲洗干净肾脏内残留的血液。与此同时，使用灌注液（附表S1）预充常温灌注环路，并将该环路加热至38℃。将肾脏的动脉套管与灌注环路的流入套管相连，使肾脏挂载于该灌注环路之上；将输尿管内的插管连接至集尿袋，以实现尿液引流。随后打开动脉流入套管，启动肾脏灌注，灌注建立后可观察到肾脏恢复粉红色外观。