A simplified subnormothermic machine perfusion system restores ischemically damaged liver grafts in a rat model of orthotopic liver transplantation
Liver donor shortages stimulate the development of strategies that incorporate damaged organs into the donor pool. Herein we present a simplified machine perfusion system without the need for oxygen carriers or temperature control, which we validated in a model of orthotopic liver transplantation.
Rat livers were procured and subnormothermically perfused with supplemented Williams E medium for 3 hours, then transplanted into healthy recipients (Fresh-SNMP group). Outcome was compared with static cold stored organs (UW-Control group). In addition, a rat liver model of donation after cardiac death was adapted using a 60-minute warm ischemic period, after which the grafts were either transplanted directly (WI group) or subnormothermically perfused and transplanted (WI-SNMP group).
One-month survival was 100% in the Fresh-SNMP and UW-Control groups, 83.3% in the WI-SNMP group and 0% in the WI group. Clinical parameters, postoperative blood work and histology did not differ significantly between survivors.
This work demonstrates for the first time in an orthotopic transplantation model that ischemically damaged livers can be regenerated effectively using practical subnormothermic machine perfusion without oxygen carriers.
- Human Resources and Services AdministrationOrgan Procurement and Transplantation Network and the Scientific Registry of Transplant Recipients: 2009 OPTN/SRTR Annual Report: Transplant Data 1999–2008 [http://optn.transplant.hrsa.gov/ar2009/]
- Schleifer D: A simple heart-lung machine for the perfusion of small laboratory animals and for organ perfusion] [in German. Chirurg 1967, 38:477–480.
- Guarrera JV, Henry SD, Samstein B, Odeh-Ramadan R, Kinkhabwala M, Goldstein MJ, Ratner LE, Renz JF, Lee HT, Brown RS Jr, Emond JC: Hypothermic machine preservation in human liver transplantation: the first clinical series. Am J Transplant 2010, 10:372–381. CrossRef
- Xu H, Lee CY, Clemens MG, Zhang JX: Prolonged hypothermic machine perfusion preserves hepatocellular function but potentiates endothelial cell dysfunction in rat livers. Transplantation 2004, 77:1676–1682. CrossRef
- Lee CY, Jain S, Duncan HM, Zhang JX, Jones JW Jr, Southard JH, Clemens MG: Survival transplantation of preserved non-heart-beating donor rat livers: preservation by hypothermic machine perfusion. Transplantation 2003, 76:1432–1436. CrossRef
- Lee CY, Zhang JX, Jones JW Jr, Southard JH, Clemens MG: Functional recovery of preserved livers following warm ischemia: improvement by machine perfusion preservation. Transplantation 2002, 74:944–951. CrossRef
- Jain S, Xu H, Duncan H, Jones JW Jr, Zhang JX, Clemens MG, Lee CY: Ex-vivo study of flow dynamics and endothelial cell structure during extended hypothermic machine perfusion preservation of livers. Cryobiology 2004, 48:322–332. CrossRef
- Saad S, Minor T: Short-term resuscitation of predamaged donor livers by brief machine perfusion: the influence of temperature. Transplant Proc 2008, 40:3321–3326. CrossRef
- Taylor MJ, Baicu S, Leman B, Greene E, Vazquez A, Brassil J: Twenty-four hour hypothermic machine perfusion preservation of porcine pancreas facilitates processing for islet isolation. Transplant Proc 2008, 40:480–482. CrossRef
- Jain S, Lee SH, Korneszczuk K, Culberson CR, Southard JH, Berthiaume F, Zhang JX, Clemens MG, Lee CY: Improved preservation of warm ischemic livers by hypothermic machine perfusion with supplemented University of Wisconsin solution. J Invest Surg 2008, 21:83–91. CrossRef
- Vogel T, Brockmann JG, Friend PJ: Ex-vivo normothermic liver perfusion: an update. Curr Opin Organ Transplant 2011, 15:167–172. CrossRef
- Perkins JD: Defatting the fatty liver with normothermic perfusion of the liver allograft. Liver Transpl 2009, 15:1366–1367.
- Brockmann J, Reddy S, Coussios C, Pigott D, Guirriero D, Hughes D, Morovat A, Roy D, Winter L, Friend PJ: Normothermic perfusion: a new paradigm for organ preservation. Ann Surg 2009, 250:1–6. CrossRef
- Reddy SP, Brockmann J, Friend PJ: Normothermic perfusion: a mini-review. Transplantation 2009, 87:631–632. CrossRef
- Mann CD, Metcalfe MS, Nicholson ML: Normothermic perfusion of ischaemic porcine kidneys: an evaluation of ex vivo function and endothelin receptor antagonism. J Nephrol 2009, 22:144–151.
- Tolboom H, Pouw RE, Izamis ML, Milwid JM, Sharma N, Soto-Gutierrez A, Nahmias Y, Uygun K, Berthiaume F, Yarmush ML: Recovery of warm ischemic rat liver grafts by normothermic extracorporeal perfusion. Transplantation 2009, 87:170–177. CrossRef
- Tolboom H, Milwid JM, Izamis ML, Uygun K, Berthiaume F, Yarmush ML: Sequential cold storage and normothermic perfusion of the ischemic rat liver. Transplant Proc 2008, 40:1306–1309. CrossRef
- Uygun K, Tolboom H, Izamis ML, Uygun B, Sharma N, Yagi H, Soto-Gutierrez A, Hertl M, Berthiaume F, Yarmush ML: Diluted blood reperfusion as a model for transplantation of ischemic rat livers: alanine aminotransferase is a direct indicator of viability. Transplant Proc 2010, 42:2463–2467. CrossRef
- Uemura R, Uchiyama K, Ozawa S, Yamaue H: Effect of normothermic perfusion using fructose-1,6-bisphosphate for maintenance of liver function duringin situextended hepatectomy by the total hepatic vascular exclusion technique. J Surg Res 2007, 137:89–95. CrossRef
- Imber CJ, St Peter SD, Lopez de Cenarruzabeitia I, Pigott D, James T, Taylor R, McGuire J, Hughes D, Butler A, Rees M, Friend PJ: Advantages of normothermic perfusion over cold storage in liver preservation. Transplantation 2002, 73:701–709. CrossRef
- Schön MR, Kollmar O, Wolf S, Schrem H, Matthes M, Akkoc N, Schnoy NC, Neuhaus P: Liver transplantation after organ preservation with normothermic extracorporeal perfusion. Ann Surg 2001, 233:114–123. CrossRef
- Kobayashi J, Ohwada S, Takeyoshi I, Ohya T, Tomizawa N, Kamoshita N, Kawashima Y, Matsumoto K, Morishita Y: Normothermic perfusion using diluted blood ameliorates ischemia-reperfusion injury on the canine liver. Transplant Proc 1998, 30:3761–3762. CrossRef
- Hellinger A, Fiegen R, Lange R, Rauen U, Schmidt U, Hirche H, Kaiser S, de Groot H, Erhard J, Eigler FW: Preservation of pig liver allografts after warm ischemia: normothermic perfusion versus cold storage. Langenbecks Arch Chir 1997, 382:175–184.
- Filipponi F, Bacci S, Romagnoli P: Normothermic liver perfusion ex situ: a resuscitation tool for hepatic grafts damaged by warm ischemia. G Chir 1993, 14:254–258.
- Bond M, Pitt M, Akoh J, Moxham T, Hoyle M, Anderson R: The effectiveness and cost-effectiveness of methods of storing donated kidneys from deceased donors: a systematic review and economic model. Health Technol Assess 2009, 13:iii-iv, xi-xiv, 1–156.
- Belzer FO, Southard JH: The future of kidney preservation. Transplantation 1980, 30:161–165. CrossRef
- Matias JE, Morais FA, Kato DM, Koziak V, Brioschi ML, Tambara EM, Agulham MÂ, Coelho JC: Prevention of normothermic hepatic ischemia during in situ liver perfusion with three different preservation solutions: experimental analysis by realtime infrared radiation thermography] [in Portuguese. Rev Col Bras Cir 2010, 37:211–217. CrossRef
- Plauth M, Zimmermann B, Raible A, Vieillard-Baron D, Bauder-Gross D, Hartmann F: Use of an artificial oxygen carrier in isolated rat liver perfusion: first demonstration of net glucose uptake at physiological portal glucose concentrations using a hemoglobin-free perfusate. Res Exp Med (Berl) 1991, 191:339–347. CrossRef
- Vairetti M, Ferrigno A, Carlucci F, Tabucchi A, Rizzo V, Boncompagni E, Neri D, Gringeri E, Freitas I, Cillo U: Subnormothermic machine perfusion protects steatotic livers against preservation injury: a potential for donor pool increase? Liver Transpl 2009, 15:20–29. CrossRef
- Vairetti M, Ferrigno A, Rizzo V, Boncompagni E, Carraro A, Gringeri E, Milanesi G, Barni S, Freitas I, Cillo U: Correlation between the liver temperature employed during machine perfusion and reperfusion damage: role of Ca 2+ . Liver Transpl 2008, 14:494–503. CrossRef
- Gringeri E, Polacco M, D’Amico FE, Scopelliti M, Bassi D, Bonsignore P, Luisetto R, Lodo E, Carraro A, Zanus G, Cillo U: A new liver autotransplantation technique using subnormothermic machine perfusion for organ preservation in a porcine model. Transplant Proc 2011, 43:997–1000. CrossRef
- Tolboom H, Izamis ML, Sharma N, Milwid JM, Uygun B, Berthiaume F, Uygun K, Yarmush ML: Subnormothermic machine perfusion at both 20°C and 30°C recovers ischemic rat livers for successful transplantation. J Surg Res in press
- Muriel A, López V, Zamora Vincente J, Gutiérrez C, Abraira Santos V, Hernández Marrero D: [Does hypothermic machine perfusion provide an advantage over cold storage in the incidence rate of delayed graft function following a deceased-donor kidney transplant?] [in Spanish]. Nefrologia 2009,29(6 Suppl):85–87.
- Delrivière L, Gibbs P, Kobayashi E, Goto S, Kamada N, Gianello P: Technical details for safer venous and biliary anastomoses for liver transplantation in the rat. Microsurgery 1998, 18:12–18. CrossRef
- Dutkowski P, Furrer K, Tian Y, Graf R, Clavien PA: Novel short-term hypothermic oxygenated perfusion (HOPE) system prevents injury in rat liver graft from non-heart beating donor. Ann Surg 2006, 244:968–977. CrossRef
- Tolboom H, Pouw R, Uygun K, Tanimura Y, Izamis ML, Berthiaume F, Yarmush ML: A model for normothermic preservation of the rat liver. Tissue Eng 2007, 13:2143–2151. CrossRef
- Kamada N, Calne RY: Orthotopic liver transplantation in the rat: technique using cuff for portal vein anastomosis and biliary drainage. Transplantation 1979, 28:47–50. CrossRef
- Mabuchi A, Wake K, Marlini M, Watanabe H, Wheatley AM: Protection by glycyrrhizin against warm ischemia-reperfusion-induced cellular injury and derangement of the microcirculatory blood flow in the rat liver. Microcirculation 2009, 16:364–376. CrossRef
- Szijártó A: Methods of increasing ischemic tolerance in liver surgery] [in Hungarian. Magy Seb 2008, 61:128–135. CrossRef
- Izamis ML, Sharma NS, Uygun B, Bieganski R, Saeidi N, Nahmias Y, Uygun K, Yarmush ML, Berthiaume F: In situ metabolic flux analysis to quantify the liver metabolic response to experimental burn injury. Biotechnol Bioeng 2011, 108:839–852. CrossRef
- Olschewski P, Gass P, Ariyakhagorn V, Jasse K, Hunold G, Menzel M, Schöning W, Schmitz V, Neuhaus P, Puhl G: The influence of storage temperature during machine perfusion on preservation quality of marginal donor livers. Cryobiology 2011, 60:337–343. CrossRef
- Sumimoto K, Inagaki K, Yamada K, Kawasaki T, Dohi K: Reliable indices for the determination of viability of grafted liver immediately after orthotopic transplantation: bile flow rate and cellular adenosine triphosphate level. Transplantation 1988, 46:506–509. CrossRef
- García-Valdecasas JC, Fondevila C: In-vivo normothermic recirculation: an update. Curr Opin Organ Transplant 2010, 15:173–176. CrossRef
- Monbaliu D, Vekemans K, De Vos R, Brassil J, Heedfeld V, Qiang L, D’Hollander M, Roskams T, Pirenne J: Hemodynamic, biochemical, and morphological characteristics during preservation of normal porcine livers by hypothermic machine perfusion. Transplant Proc 2007, 39:2652–2658. CrossRef
- Radunz S, Paul A, Nowak K, Treckmann JW, Saner FH, Mathé Z: Liver transplantation using donor organs with markedly elevated liver enzymes: how far can we go? Liver Int 2011, 31:1021–1027. CrossRef
- Monbaliu D, Pirenne J, Talbot D: Liver transplantation using donation after cardiac death donors. J Hepatol 2012, 56:474–485. CrossRef
- A simplified subnormothermic machine perfusion system restores ischemically damaged liver grafts in a rat model of orthotopic liver transplantation
- Open Access
- Available under Open Access This content is freely available online to anyone, anywhere at any time.
- Online Date
- May 2012
- Online ISSN
- BioMed Central
- Additional Links
- Author Affiliations
- 1. Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, 51 Blossom Street, Boston, MA, 02114, USA
- 2. Department of Biomedical Engineering, Rutgers University, 599 Taylor Road, Piscataway, NJ, 08854, USA