Orthotopic liver transplantation following dual machine perfusion in the mouse

M. Fujiyoshi, T.A. Berendsen, R. van Rijn, R.J. Porte

Chair(s): dr. Martin. J. Hoogduijn, wetenschappelijk medewerker, Erasmus MC & prof. dr. Cees. van Kooten, onderzoeker, LUMC

Wednesday 9 march 2016

15:50 - 16:00h at Zaal 5 & 6

Categories: Parallelsessie (basaal)

Parallel session: Parallelsessie IV - Basaal

Ex vivo Machine perfusion of explanted organs can improve the utilization of grafts derived from extended criteria donors and donors after circulatory death. To effectively study and improve clinical machine perfusion, we developed a murine model of hypothermic dual machine perfusion and orthotopic liver transplantation. This model enables a host of benefits ranging from high-throughput screening of pharaceutical enhancements to using knockout technology to elucidate fundamental pathological processes during liver transplantation.

Machine perfusion: The perfusion machine comprised of two independent perfusion circuits for portal venous and arterial perfusion and a perfusate temperature control system. We harvested livers from five Male C57BL/6 mice aged 9–12 weeks, preserving the hepatic arteries (HA), superior mesenteric artery (SMA), and abdominal aorta, preserving sufficient length of the supra-hepatic vena cava (SHVC), infra-hepatic vena cava (IHVC), and portal vein (PV). After vascular cuffs were attached to the IHVC and PV, the graft was connected to the portal and venous perfusion port, respectively. Dual machine perfusion was performed at 4oC for 30 minutes using William's medium E based perfusion solution. The PV flow rate was set at 4ml/min, the arterial flow rate at 0.5ml/min. Liver transplantation: The recipient procedure (n=5) was started after 30 minutes of MP. Upon graft introduction into the recipient abdomen, portal blood flow was established first, after which the IHVC was anastomosed using a cuff technique. Arterial reconstruction was then performed using side-to-end anastomosis between the abdominal aorta of the graft and recipient. The bile duct was reconstructed using an intraluminal stent.

During hypothermic dual machine perfusion, all perfusion parameters remained within normal range. The mean total perfusion time was 82 minutes. All recipient mice recovered from anesthesia after the operation. The 3-day survival rate was 80% (4/5).

Herein we established a novel murine model of hypothermic dual machine perfusion, which was confirmed with orthotopic liver transplantation. This model is suitable for a wide range of both fundamental and clinically applied research, which will be vital to optimizing the benefits of machine perfusion.