急性肝功能衰竭猪应用同种体外肝脏灌注技术支持的实验研究

目的 评价体外肝脏灌注（extracorporeal liver perfusion,ECLP）技术应用于急性肝功能衰竭（acute liver failure,ALF）短暂替代治疗的疗效和可行性。方法 实验动物均为健康普通长白猪,体重20～30kg。按随机数字表法随机分为3组：肝衰组（n=5）,通过结扎肝脏血供和门体分流制备肝衰模型,肝衰模型制成8h后处死取标本;肝衰＋ECLP组（n=5）,受体为肝衰猪,供肝阻断血供后迅速切取,连接ECLP开始灌注,ECLP灌注时间为4h,肝衰模型制成8h后处死取标本;正常肝＋ECLP组（n=4）,受体为正常猪,灌注方法同肝衰＋ECLP组。观察受体一般情况、肝脏和脑组织病理变化,检测受体血常规、血生化、血凝、血氨和TNF等指标。结果 肝衰组PT、AST、TNF、血氨、RBC和HCT值明显高于肝衰＋ECLP组（P〈0．05）;正常肝＋ECLP组的FIB、AST、TNF和血氨的变化明显低于肝衰＋ECLP组（P〈0．05）。病理学检查显示,肝衰组受体的脑组织出现广泛的脑水肿,表现为神经元细胞问隙明显增宽,并有多数神经元细胞死亡;肝衰＋ECLP组受体的脑组织也出现有脑水肿的表现,并有少数神经元细胞死亡,但较肝衰组轻;正常肝＋ECLP组的脑组织病理检查基本正常。肝衰组和肝衰＋ECLP组的受体肝脏病理检查可见大片肝细胞坏死,而正常肝＋ECLP组受体的肝脏病理检查基本正常。结论 同种ECLP能有效地改善肝衰受体的体内环境,缓解症状．尤其是这种技术能缓解肝衰受体的脑水肿,而脑水肿可能是肝衰受体最主要的致死原因,但ECLP也有其自身的并发症存在。总之,同种ECLP技术是一种有效而且可行的ALF短暂替代治疗方案。     

体外持续肝脏灌注常温保存和HTK液低温保存无心跳供肝效果的比较

目的 比较应用组氨酸-色氨酸-酮戊二酸(HTK)液低温保存和体外持续肝脏灌注(ECLP)系统常温保存无心跳供肝的效果.方法 按保存方法不同将供肝随机分为A组和B组:供肝切取后,A组用HTK液在低温下保存10 h;B组用ECLP系统在常温下用稀释的自体血液持续灌注10 h.两组供肝再经过60 min冷缺血期后,连接ECLP系统用稀释的自体血液再灌注4 h.观察再灌注后1、2、3、4 h四个时间点的胆汁分泌量,门静脉和肝动脉的压力,肝脏耗氧率的变化,灌注液中丙氨酸转氨酶(ALT)、乳酸脱氢酶(LDH)、葡萄糖水平以及灌注后供肝的常规病理和超微病理变化.结果 B组再灌注后1、2、3、4 h时间点的胆汁分泌量,门静脉和肝动脉的压力,灌注液中ALT、LDH和葡萄糖水平,以及2、3、4 h时间点的耗氧率与A组比较,差异均有统计学意义(P＜0.05或P＜0.01);B组供肝的病理损害程度较A组轻.结论 供肝切取后10 h内,利用ECLP系统持续灌注常温保存比用HTK液单纯低温保存在维持无心跳供肝的功能和生理活性方面效果更好.     

丹参对肝脏再灌注损伤的防护作用

丹参在防止肝脏缺血再灌注损伤中的作用日益受到人们的重视[1,2 ] 。为了解丹参对移植肝再灌注损伤的防护作用 ,我们用含丹参的乳酸林格液直接灌洗保存猪肝 ,并将保存后的肝脏进行移植。一、材料与方法1.动物和分组 :杂交长白猪 76头 ,雌雄不拘 ,体重 18～ 32ｋｇ ,随机分为供肝组和移植组 ,每组 38头。共施行肝移植术 38次 ,其中辅助性肝移植 2 6次 ,背驮式原位肝移植 12次。将移植组的 38头猪随机分为 3组 :Ａ组 (对照组 ) 11头 ,Ｂ组 (静脉滴注丹参组 ) 12头 ,Ｃ组(丹参灌洗保存肝脏 静脉滴注丹参组 )15头。2 .实验方法 :供肝切取均采用…     

体外肝脏切除术对肝脏功能和结构的影响

方法:正常家猪13头，在体外静脉转流和4℃EuroCollins液持续灌注肝脏下行体外肝切除及自体残肝原位再植术。在手术中不同时相点检测肝组织能荷水平（EC）、线粒体呼吸活性（RCR）、动脉血酮体比值（AKBR）和肝脏病理变化。结果：在肝脏冷灌注时相，两组动物肝组织ATP、EC及AKBR水平较术前显著降低，RCR轻度降低。残肝植入后，术程平稳的6头动物，EC水平回升、RCR无显著变化、AKBR回升到0．7以上；术程不平稳的7头动物，RCR下降、EC进一步降低、AKBR仍低于0．5。冷灌注时肝脏组织结构基本保持完整，残肝再植复流后肝组织出现以肝腺泡第Ⅲ带为著的组织损害。结论：体外肝切除术造成肝脏损害的主要原因是再灌流损害；肝细胞能量代谢状态与手术动物的预后密切相关。     

hDAF转基因猪在肝脏离体灌注实验中对超急性排斥反应的防止作用

目的 在离体肝脏灌注实验中,观察人衰变加速因子（hDAF)转基因猪对肝脏超急性排斥反应的阻止作用。方法 全麻下分别分离出门静脉、肝上及肝下下腔静脉、腹主动脉,分别经腹主动脉和门静脉插管,用UW液对肝脏进行原位灌注,然后摘取肝脏置于4℃的冷容器中,冷缺血时间少于1h;灌注中,血液经肝下下腔静脉流出,循经膜氧和器一热交换器,然后经门静脉返回肝脏,血流速度0.75-1ml/克肝组织,共灌注16只猪,将其分为猪血灌注猪肝脏组（P-P组,n=6)、人血灌注猪肝脏组（H-P组,n=5)和人血灌注hDAF转基因猪肝脏组（H-hDAF组,n=5)。结果 经组织学检查,P-P组有正常的组织学结构;H-P组显示有明显的血管内皮细胞的损害、出血、血小板及纤维蛋白的沉积;H-hDAF组除1例有轻度的血管内皮损害和出血外,其余为基本正常的组织学结构。H-hDAF组在灌注过程中的胆汁产量,凝血功能及门脉压力的变化都明显好于H-P组。结论 hDAF的转基因猪对肝脏的超急性排斥反应有明显的阻止作用,这为将来临床应用hDAF的转基因猪肝脏做体外灌注支持治疗提供了理论支持。     

小鼠肝脏体外复灌系统的建立

目的建立小鼠肝脏体外复灌系统，并验证其有效性。 方法6只雄性C57小鼠获取肝脏后冷保存45 min内行体外复灌，复灌前获取同种属小鼠血液配制复灌液。小鼠复灌系统以多轴蠕动泵作为动力，实行门静脉单通道灌注，在灌注系统中加入输液器滴斗以缓冲压力和排气，2个小动物膜式氧合器串联以达到携氧要求，灌注系统总液体量约30 mL，复灌流量设定为2.5 mL·min^－1·g^－1。采用双托盘形式，满足小鼠肝脏正常体位。应用生物机能实验系统记录入肝压力等灌注参数，监测灌注液ALT、AST、肝组织病理及细胞凋亡等指标。 结果小鼠肝脏体外复灌过程中，门静脉压力低于8 mmHg，系统压力波动小于1 mmHg，灌注液pH值维持在7.40±0.05。复灌6 h结束时，灌注液ALT、AST和乳酸脱氢酶分别为(62±14)、(160±23)和(682±71) U/L，乳酸为(3.2±0.1) mmol/L。肝组织病理示肝小叶结构完整，实质细胞无明显坏死，病理学评分(1.1±0.7)分。原位缺口末端标记法检测肝细胞凋亡率为(0.57±0.22)%。 结论本研究国内首次构建了小鼠肝脏体外复灌系统，操作简单、运行稳定，能够较长时间维持肝脏正常生理功能。     

常温局部灌注在受控心脏死亡肝移植供体中的应用

目前全球器官移植领域所面临的仍是供体短缺的主要问题。由于受体需求量和标准供体的数量的严重不平衡，外科医生们将目光转移到心脏死亡供体（DCD）。相比于脑死亡供体（DBD），DCD面临的主要问题是经历更长的热缺血时间（WIT），以致患者术后并发症发生率增加，尤其是胆道并发症。受控心脏死亡供体（cDCD）是指在符合患者或家属意愿的情况下有计划地退出维持生命的治疗，在一段“无接触”时间后（通常为5 min），宣布患者大脑循环永久缺失，同时快速进行器官恢复。由于使用常规器官保存方法保存的DCD移植物的移植效果不尽如人意，近年来，常温灌注技术所展现出的优势在移植过程中愈发明显，逐渐引起外科医生和科学家们的关注。与活体肝移植以及DBD肝移植不同，DCD在宣布死亡前无法取出移植物。而在常温局部灌注（NRP）中，外科医生们能够在宣布死亡后阻断流向大脑的血液，并通过体外膜氧合启动器官的热灌注，在供体体内恢复供体肝脏的功能，使肝脏产生胆汁并清除乳酸。这一过程为供体肝脏离开供体和移植前的各项指标监测和优化提供宝贵时间。目前已有多项临床研究表明，NRP作为一种原位器官修复技术能够使cDCD供体肝脏的移植效果与DBD供体相近。另外，一些研究者还开发了多种灌注技术的联合应用，包括NRP与机器灌注（MP）以及双低温氧合机灌注（D-HOPE）的联合使用，均展现出良好的移植效果，为肝移植供体保存提供更多可能性。虽然许多学者认为NRP是一种获取更多高质量器官的保存方法，但有研究者质疑该技术的伦理问题。他们认为该技术违背死亡捐赠规则，NRP灌注程序中所涉及的操作可能导致患者的死亡，因此，确保患者的循环以及呼吸的永久性不可恢复状态以及在NRP期间确保脑部循环的缺失尤其重要。鉴于NRP在移植领域的重要性，笔者对NRP技术在cDCD中的应用进行归纳总结。     

犬辅助性原位肝左叶移植模型的建立

目的建立稳定可靠的辅助性肝移植动物模型。方法杂交犬14条8～25kg随机分两组。供体组肝脏均采用尸体肝左叶；受体组行标准左叶切除，供肝左叶原位移植于受体体内。结果供肝热缺血时间为零，冷缺血时间平均36．3min；灌注液的量平均2．96L，切取修剪肝左叶时间为23～40min。受体手术平均时间5．3h，平均出血量140ml，肝脏血管重建后红润柔软，5～11min内即见胆汁从胆管中溢出。受体组术后全部存活，围手术期未用任何血管活性药物、抗生素及免疫抑制剂。术后存活超过6h者5例．最长存活者达5．7d。结论犬是建立辅助性肝移植模型的理想动物。     

半离体体外肝手术不同术式对肝脏缺血再灌注损伤的影响

目的 观察3种半离体体外肝手术方式对肝脏缺血再灌注损伤的影响。方法 SD大鼠随机分为对照组（A组）、单纯离断肝下下腔静脉手术组（B组）、单纯离断肝上下腔静脉手术组（C组）和联合离断肝上、肝下下腔静脉手术组（D组），每组各10只，手术后4、24h分组收集血清和肝组织，分别进行血清谷丙转氨酶（ALT）监测、病理切片、免疫组织化学和逆转录-聚合酶链反应（RT—PCR）检测。结果 ALT随各组手术复杂程度和再灌注时间的增加而升高；病理切片见各手术组肝脏炎症反应随手术复杂程度和再灌注时间的增加而有所加重；B组术后4和24h肝组织中过氧化物酶体增殖激活受体（PPAR）α的表达无明显变化，但术后24h原癌基因B淋巴瘤-xL（bcl-xL）的表达最强；C、D组肝组织术后4hPPARα和bcl—xL的表达增加，术后24hPPARα的表达明显强于4h组．而bcl—xL的表达较4h明显下降。结论 PPARα和bcl—xL参与体外肝手术后肝组织缺血再灌注损伤，两者的表达在一定程度上反应了肝细胞受损的程度。B组手术方式简单、对肝脏的损伤较小；C、D两组手术复杂，对肝组织的再灌注损伤较重。     

丹参对大鼠肝脏低温保存损伤的保护作用

目的　研究丹参对大鼠肝脏低温保存损伤的保护作用。方法　采用大鼠肝脏离体非循环灌注模型 ,观察乳酸林格液 (LR液 )中加入不同剂量丹参后保存大鼠肝脏 1 2h的效果。结果　丹参组 ( 60 0mg/g)肝组织ATP含量 ( 6.0 8± 0 .67) μmol/g及分泌胆汁量 ( 1 0 5.6± 1 2 .4) μl/h明显高于对照组 ( 2 .52± 0 .31 ) μmol/g及 ( 57.4± 8.2 ) μl/h(P <0 .0 5) ,丹参组 ( 60 0mg/g)肝组织 2 ,3 二羟苯甲酸 ( 2 ,3 DHBA) ( 0 .1 54± 0 .0 1 3)nmol/g、2 ,5 DHBA( 1 .354± 0 .0 68)nmol/g及流出液天门冬氨酸转氨酶 (AST) ( 38.4± 3.7)U/L明显低于对照组 ( 0 .2 4 5± 0 .0 2 1 )nmol/g、( 2 .1 0 5± 0 .0 97)nmol/g及( 76.4± 9.2 )U/L ,差异有显著性 (P <0 .0 5)。结论　丹参可明显提高供体肝脏的保存效果 ,其作用机理可能主要与改善低温保存肝脏的能量代谢及抗氧自由基的作用有关     

常温机械灌注修复长时间冷保存供肝的大动物实验研究

目的探索常温机械灌注(NMP)在挽救大动物边缘供肝中的价值。 方法6只雄性、10～12月龄巴马小型猪分为静态冷保存(SCS)6 h组和SCS 24 h组,每组3只,分别于供肝获取后SCS 6 h、24 h后进行2 h NMP复苏。基于荷兰Organ Assist公司Liver Assist系统和供体猪自身血液,整合四通道生理仪器及自制灌注管路搭建NMP平台。在NMP过程中分别于灌注0、15、60、90、120 min 5个时间点收集灌注液用于肝功能(ALT、AST)检测及血气分析(pH、氧分压)。灌注结束取肝左叶相同位置少许组织,以10%甲醛固定,用于后续HE染色。 结果NMP开始时,SCS 24 h组ALT、AST水平略高于SCS 6 h组,之后AST、ALT水平均缓慢上升。NMP开始时SCS 24 h组pH值中位数为7.28,氧分压中位数为46 mmHg(1 mmHg＝0.133 kPa,下同),均低于SCS 6 h组(7.36,52 mmHg),经过2 h NMP后SCS 24 h组pH值和氧分压逐渐接近甚至优于SCS 6 h组。HE染色发现,灌注前SCS 24 h组肝脏充血严重,炎性细胞浸润明显;NMP 2 h后,肝脏充血明显改善、炎性细胞减少,与SCS 6 h组无明显差异。 结论基于Liver Assist系统和供体猪自身血液成功搭建的NMP平台,可有效改善长时间冷保存的边缘供肝质量,提示NMP在修复边缘供肝、扩展供肝来源方面具有重要临床及基础研究价值。     

HDAF transgenic pig livers are protected from hyperacute rejection during ex vivo perfusion with human blood

The aim of this study was to determine if human decay-accelerating factor (hDAF) protects against hyperacute rejection in an ex vivo liver perfusion system using human blood. Pig livers were perfused ex vivo via the portal vein for an average of 5–6 h using a membrane oxygenator. Three groups were studied. Group I: Wild-type pig livers were alloperfused with fresh pig blood (n=5). Group II: Wild-type pig livers were xenoperfused with fresh human blood (n=5). Group III: hDAF transgenic pig livers were xenoperfused with fresh human blood (n=5). The graft ischemic time, ratio of perfusate volume to liver weight, flow rate, and perfusate hematocrit were similar in each group. The hDAF livers perfused with human blood (Group III) had a lower ALT level, less protein and albumin losses, lower bilirubin levels in the perfusate, less weight gain, and greater bile production than the wild-type livers perfused with human blood. Histology showed classic features of hyperacute rejection in Group II, including massive hemorrhage, severe vasculitits, fibrin and C5b-9 deposition, and endothelial damage within 1 h of perfusion, whereas liver histology studies in Groups I and III were near normal. IgG and IgM deposits were seen in the xenoperfused livers. Electron microscopy (EM) and immuno-EM showed loss of endothelial cells, trapping of white blood cells and platelets, and diffuse fibrin deposits in Group II only. hDAF pig livers perfused with human blood showed superior function and histology when compared with wild-type pig livers. These data suggest that (1) hyperacute rejection may contribute to the inconsistent results using wild-type pig livers for extracorporeal liver support and (2) genetically modified pigs that express hDAF may provide a better donor source than wild-type pigs for extracorporeal liver support.     

Assessment of the AMC-bioartificial liver in the anhepatic pig

BACKGROUND: The anhepatic pig model was used to evaluate a bioartificial liver developed in our institution (AMC-BAL). The bioartificial liver is based on oxygenated plasma perfusion of porcine hepatocytes attached to a polyester matrix. METHODS: Pigs (n=15) underwent total hepatectomy with restoration of caval continuity using a polyethylene, three-way prosthesis. In group I, pigs received limited intensive care under continuation of general anesthesia (n=5). Group II pigs (n=5) underwent, in addition, extracorporeal plasma perfusion of an AMC-BAL without hepatocytes (device control group). In group III (n=5), plasma perfusion occurred with an AMC-BAL loaded with autologous hepatocytes. Groups II and III were connected to the extracorporeal system 24 hr after hepatectomy, for a period of 24 hr. The main outcome parameters were as follows: survival time, liver enzymes (aspartate aminotransferase, alanine aminotransferase), blood ammonia, and total/direct bilirubin. RESULTS: Survival (mean +/- SD) of the anhepatic pigs was significantly increased in the BAL-treated group (group III: 65+/-15 hr), as compared with the control groups (group I: 46+/-6 hr and group II: 43+/-14 hr). Mean blood ammonia levels during BAL treatment were significantly lower in the BAL-treated group in comparison with both control groups (P=0.02). Total and direct bilirubin levels gradually increased after hepatectomy and reached maximum values of 1.98 mg/dl and 1.50 mg/dl, respectively, showing no differences between the three groups. CONCLUSIONS: (1) Treatment of anhepatic pigs with the AMC-BAL containing autologous hepatocytes significantly increases survival time, which is associated with a significant decrease in blood ammonia. 2) Anhepatic pigs demonstrate increasing direct bilirubin levels as a result of extrahepatic bilirubin conjugation.     

Isolated hyperthermic liver perfusion with high dose tumor necrosis factor alpha in pigs: an experimental study in preparation of clinical Use

Isolated hyperthermic perfusion of the liver was performed for 45 min in 27 pigs via hepatic artery and portal vein at mean inflow temperatures between 40.7 and 41.2 degrees C. In two study groups B and C (n = 9 pigs each) 50 microg recombinant human tumor necrosis factor-alpha (rhTNFalpha) per kg body weight were added to the perfusate, whereas in a control group A liver perfusion was done without rhTNFalpha. Before reperfusion the livers were washed out with Ringer's solution in all groups followed by a protein solution in group C. At 30 and 60 min after reperfusion the maximum systemic rhTNFalpha concentrations were significantly higher in group B with 68 and 61 ng/ml compared to 14.5 and 14.9 ng/ml in group C (p < 0. 05). Mean systemic porcine TNFalpha concentration was significantly higher in group B (217 pg/ml) compared to group C (50 pg/ml) 30 min after reperfusion (p = 0.012). Survival was 7/9 in group A and C and only 2/9 in group B with 6/7 pigs dying due to severe cardiopulmonary failure within 12 h after operation. In surviving pigs of group A and C only mild and transient hepatotoxicity was registered. The presented study underlines the feasibility of high dose rhTNFalpha application in an isolated hyperthermic liver perfusion system. Washout of the liver with a protein solution before reperfusion reduces systemic TNFalpha levels as well as associated lethal cardiocirculatory and hepatotoxic side effects.     

Extended preservation of non-heart-beating donor livers with normothermic machine perfusion

BACKGROUND: Non-heart-beating donor (NHBD) livers represent an important organ pool, but are seldom utilized clinically and require rapid retrieval and implantation. Experimental work with oxygenated perfusion during preservation has shown promising results by recovering function in these livers. This study compared sanguinous perfusion with cold storage for extended preservation of the NHBD liver in a porcine model. METHODS: Porcine livers were subjected to 60 min of in vivo total warm ischaemia before flushing, after which they were preserved by one of two methods: group 1 (n = 4), University of Wisconsin (UW) solution by standard cold storage for 24 h; group 2 (n = 4), oxygenated autologous blood perfusion on an extracorporeal circuit for 24 h. All livers were subsequently tested on the circuit during a 24-h reperfusion phase. RESULTS: Livers in group 1 showed no evidence of viability during the reperfusion phase with no bile production or glucose utilization; they also displayed massive necrosis. Livers in group 2 demonstrated recovery of function by synthetic function, substrate utilization and perfusion haemodynamics; these livers displayed less cellular injury by hepatocellular enzymes. All differences in parameters between the two groups were statistically significant (P < 0.05). These findings were supported by histological examination. CONCLUSION: Warm ischaemia for 1 h and simple cold storage (UW solution) for 24 h renders the liver non-viable. Oxygenated, sanguinous perfusion as a method of preservation recovers liver function to a viable level after 24 h of preservation.     

Comparative study of bioartificial liver support and plasma exchange for treatment of pigs with fulminant hepatic failure

Recently, bioartificial liver (BAL) treatment was reported to provide beneficial effects for patients with fulminant hepatic failure (FHF). Some success in experimental or clinical trials has been reported; however, the evaluation of BAL efficacy remains unclear, especially in comparison with other treatments for FHF. The purpose of this study was to compare the efficacy between BAL and plasma exchange (PE) in experimentally induced FHF in pigs. Pigs undergoing hepatic devascularization (HD) were placed into the following groups: no treatment (control; n = 6), BAL treatment (BAL; n = 5), and plasma exchange (PE; n = 5). Each treatment was initiated 6 h after HD and lasted for 4 h. BAL treatment significantly improved liver functions in FHF pigs. The decrease in cerebral perfusion pressure was also significantly suppressed in the pigs with BAL, and their survival time was prolonged compared with the results in pigs with PE. The effects of BAL outperform those of PE in the treatment of experimental FHF model.     

Liver transplantation for acute liver failure: trying to define when transplantation is futile

BACKGROUND: Recently, the model of end-stage liver disease (MELD) index has been used to select patients with acute liver failure (ALF) or transplantation. By the time the indication for orthotopic liver transplantation (OLT) is defined, the patient's clinical status may worsen. OBJECTIVE: In this study, MELD was used to define patients beyond OLT. METHODS: Among adult patients ALF was responsible for 17 OLT. Their medical records were reviewed to calculate the MELD score just before the OLT. MELD of the deceased patients after OLT (group 1, n=8), was compared with the MELD score of living recipients (group 2, n=9). Creatinine level, need for dialysis, use of vasoactive amines, and mechanical ventilation before OLT were also analyzed in these groups. A significant difference was defined when P<.05. RESULTS: The mean MELD score+/-SD was 51.86+/-12.3 for group 1, and 38.47+/-7.1 for group 2 (P=.02). There was no difference between the creatinine values for patients in the 2 groups (P=.20). Also, the use of vasoactive amines or the need of dialysis before OLT were not different (P=.12 and P=.25, respectively). Group 1 was more frequently under mechanical ventilation, and showed a 4.29 relative risk for death after OLT. CONCLUSION: MELD score could be useful to define the prognosis of OLT among patients with ALF.     

Study of kidney and liver viability in the rat after exclusive aortic perfusion using intracellular ATP measurement

Summary To find whether the liver can be procured after exclusive aortic perfusion, three organ perfusion models were used in three groups of donor rats. Group 1 underwent liver wash-out via the portal vein; in group 2, the kidneys alone were perfused via the aorta; and group 3 underwent simultancous aortic perfusion of liver and kidneys. All perfusion flow rates in the three groups were adjusted to physiological values. Harvested organs were transplanted and recipient animals were killed 4h after transplantation to study liver and kidney viability by using intracellular ATP measurement. Liver ATP was lower (P< 0.005) in the portal perfusion group (group 1: 1.396±0.412) than in the aortic perfusion group (group 3: 2.181±0.061). Kidney ATP was comparable in groups 2 and 3: 1.066±0.09 vs 1.059±0.273 (mol/g) tissue. Liver cooling was quicker with portal perfusion than with the aortic flush (20°C in 20 s vs 15°C in 60 s). Aortic perfusion at a physiologic flow rate has no detrimental effect on renal viability studied by intracellular ATP measurement. We conclude that liver cooding via the aortic route only is a good alternative to portal perfusion and seems to give good preservation. Application of this observation to emergency procurement in humans is still the subject of controversy.