Orthotopic liver transplantation from cardiac death donors in the mouse: a new model and evaluation of cardiac death time

Document Type : Original Article


1 Department of General Surgery, The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province, China

2 Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Peking University Cancer Hospital and Institute, Beijing, China

3 Department of Anesthesiology, The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning Province, China

4 Department of General Surgery, Anhui Provincial Hospital, Hefei, Anhui, China


Objective(s): The goal of this research was to develop a mouse orthotopic liver transplantation (LTx) model from donor-after-cardiac-death (DCD) grafts.
Materials and Methods: Mice were randomly assigned to the experimental group or the sham group. The mice in the experimental group were divided into three groups according to the warm ischemia time (WIT) of liver graft: normal LTx, WIT 30 minute (min) +LTx and WIT 45 min +LTx. The descending aorta was clamped using a miniature aortic clamp to simulate cardiac arrest in the DCD grafts. Subsequently, the grafts were orthotopically transplanted into C57BL/6 mice. The 7-day survival rate, serum alanine aminotransferase (ALT), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) mRNA level, tumor necrosis factor-alpha (TNF-α) mRNA level, as well as hepatic pathologic alterations were observed.
Results: The 7-day survival rate was markedly lower in the WIT 45 min+LTx group than that in the normal LTx group (25% versus 100%, P-value<0.05), with no significant difference between the WIT 30 min +LTx and normal LTx group (75% versus 100%, P-value>0.05). Serum ALT level of WIT 45 min+LTx group was markedly higher than that of normal LTx and WIT 30 min+LTx group (P-value<0.01). There were significant differences in necrosis and apoptosis among the three groups (P-value<0.05). The expression of iNOS, IL-6 mRNA and TNF-α mRNA in WIT 45 min +LTx group all increased significantly compared with the normal LTx and WIT 30 min+LTx group.
Conclusion: The DCD LTx model is feasible in the mouse and would provide many advantages for biomedical research on LTx from DCD grafts.


1. Detry O, Donckier V, Lucidi V, Ysebaert D, Chapelle T, Lerut J, et al. Liver transplantation from donation after cardiac death donors: initial Belgian experience 2003-2007. Transpl Int 2010; 23:611-618.
2. Dubbeld J, van Hoek B, Ringers J. Use of a liver from donor after cardiac death: is it appropriate for the sick or the stable? Curr Opin Organ Transplant 2011; 16:239-242.
3. Liu Q, Rehman H, Krishnasamy Y, Ramshesh VK, Theruvath TP, Chavin KD, et al. Role of inducible nitric oxide synthase in mitochondrial depolarization and graft injury after transplantation of fatty livers. Free Radic Biol Med 2012; 53:250-259.
4. Jay CL, Skaro AI, Ladner DP, Wang E, Lyuksemburg V, Chang Y, et al. Comparative effectiveness of donation after cardiac death versus donation after brain death liver transplantation: Recognizing who can benefit. Liver Transpl 2012; 18:630-640.
5. Wertheim JA, Petrowsky H, Saab S, Kupiec-Weglinski JW, Busuttil RW. Major challenges limiting liver transplantation in the United States. Am J Transplant 2011; 11:1773-1784.
6. Tuttle-Newhall JE, Krishnan SM, Levy MF, McBride V, Orlowski JP, Sung RS. Organ donation and utilization in the United States: 1998-2007. Am J Transplant 2009; 9:879-893.
7. de Vera ME, Lopez-Solis R, Dvorchik I, Campos S, Morris W, Demetris AJ, et al. Liver transplantation using donation after cardiac death donors: long-term follow-up from a single center. Am J Transplant 2009; 9:773-781.
8. Heidenhain C, Pratschke J, Puhl G, Neumann U, Pascher A, Veltzke-Schlieker W, et al. Incidence of and risk factors for ischemic-type biliary lesions following orthotopic liver transplantation. Transplant Int 2010; 23:14-22.
9. Foley DP, Fernandez LA, Leverson G, Anderson M, Mezrich J, Sollinger HW, et al. Biliary complications after liver transplantation from donation after cardiac death donors: an analysis of risk factors and long-term outcomes from a single center. Ann Surg 2011; 253:817-825.
10. Wigmore SJ. Similar liver transplantation survival with selected cardiac death donors and brain death donors (Br J Surg 2010; 97: 744-753). Br J Surg 2010; 97:753.
11. Xia W, Ke Q, Wang Y, Feng X, Guo H, Wang W, et al. Donation after cardiac death liver transplantation: Graft quality evaluation based on pretransplant liver biopsy. Liver Transpl 2015; 21:838-846.
12. de Rougemont O, Breitenstein S, Leskosek B, Weber A, Graf R, Clavien PA, et al. One hour hypothermic oxygenated perfusion (HOPE) protects nonviable liver allografts donated after cardiac death. Ann Surg 2009; 250:674-683.
13. Kageyama S, Yagi S, Tanaka H, Saito S, Nagai K, Hata K, et al. Graft reconditioning with nitric oxide gas in rat liver transplantation from cardiac death donors. Transplantation 2014; 97:618-625.
14. Muiesan P, Fisher S. The bile duct in donation after cardiac death donor liver transplant. Curr Opin Organ Transplant 2014; 19:447-452.
15. Chen J, Gong W, Ge F, Huang T, Wu D, Liang T. A review of various techniques of mouse liver transplantation. Transplant Proc 2013; 45:2517-2521.
16. Miyagi S, Ohkohchi N, Oikawa K, Satoh M, Tsukamoto S, Satomi S. Effects of anti-inflammatory cytokine agent (FR167653) and serine protease inhibitor on warm ischemia-reperfusion injury of the liver graft. Transplantation 2004; 77:1487-1493.
17. Schlegel A, Graf R, Clavien PA, Dutkowski P. Hypothermic oxygenated perfusion (HOPE) protects from biliary injury in a rodent model of DCD liver transplantation. J Hepatol 2013; 59:984-991.
18. Bezinover D, Ramamoorthy S, Postula M, Weller G, Mahmoud S, Mani H, et al. Effect of cold perfusion and perfluorocarbons on liver graft ischemia in a donation after cardiac death model. J Surg Res 2014; 188:517-526.
19. Chen L, Chen G, Guo Y, Liu L, Xiao L, Fan W, et al. Ketanserin, a serotonin 2A receptor antagonist, alleviates ischemia-related biliary fibrosis following donation after cardiac death liver transplantation in rats. Liver Transpl 2014; 20:1317-1326.
20. Liu Q, Rehman H, Krishnasamy Y, Schnellmann RG, Lemasters JJ, Zhong Z. Improvement of liver injury and survival by JNK2 and iNOS deficiency in liver transplants from cardiac death mice. J Hepatol 2015; 63:68-74.
21. Tian Y, Rudiger HA, Jochum W, Clavien PA. Comparison of arterialized and nonarterialized orthotopic liver transplantation in mice: prowess or relevant model? Transplantation 2002; 74:1242-1246.
22. Zhou S, Palanisamy AP, McGillicuddy JW, Theruvath TP, Emre SH, Chavin KD. New method of stent-facilitated arterial reconstruction for orthotopic mouse liver transplantation. J Surg Res 2014; 187:297-301.
23. Pan N, Liu Z, He J, Li S, Lv X, Wang L, et al. Comparison of methods for the reconstruction of the hepatic artery in mouse orthotopic liver transplantation. PLoS One 2015; 10:e0133030.
24. Zhong Z, Ramshesh VK, Rehman H, Currin RT, Sridharan V, Theruvath TP, et al. Activation of the oxygen-sensing signal cascade prevents mitochondrial injury after mouse liver ischemia-reperfusion. Am J Physiol Gastrointest Liver Physiol 2008; 295:G823-832.
25. Liu Q, Rehman H, Shi Y, Krishnasamy Y, Lemasters JJ, Smith CD, et al. Inhibition of sphingosine kinase-2 suppresses inflammation and attenuates graft injury after liver transplantation in rats. PLoS One 2012; 7:e41834.
26. Karp SJ, Johnson S, Evenson A, Curry MP, Manning D, Malik R, et al. Minimising cold ischaemic time is essential in cardiac death donor-associated liver transplantation. HPB (Oxford) 2011; 13:411-416.
27. Reich DJ, Mulligan DC, Abt PL, Pruett TL, Abecassis MM, D'Alessandro A, et al. ASTS recommended practice guidelines for controlled donation after cardiac death organ procurement and transplantation. Am J Transplant 2009; 9:2004-2011.