大鼠辅助性肝移植诱导免疫耐受模型的建立

目的利用PVG（TR-1^c）和DA（RT-1^a）大鼠分别为供受体，联合进行辅助性肝移植和异位心脏移植，建立大鼠辅助性肝移植诱导免疫耐受模型。方法辅助性肝移植采用重建门静脉法，异位心脏移植采用腹腔吻合法。结果同品系对照组和实验组动物及其移植心脏存活超过100d，而异品系对照组移植心脏只存活7d。结论在受体肝脏存在的情况下，辅助性供肝依然发挥其诱导耐受的效应。经长期观察，供肝无萎缩，耐受诱导效应持续存在，指示心脏搏动良好。因此，该模型在肝移植基础研究中具有实用价值。     

The combi-effect--reduced rejection of the heart by combined transplantation with the lung or spleen.

The term combi-effect was introduced to describe the phenomenon of a reduction in rejection of heart grafts after combined transplantation with the lung. In this study in rats we investigated whether the combi-effect was an immunological process and whether it could also be induced by combined transplantation of the heart with the spleen or with a lymphocyte-depleted spleen. Heart and spleen grafts were transplanted into the abdomen; left lungs were transplanted into the thorax of recipient rats. To deplete spleens of their lymphocytes, prospective donor rats were irradiated. Cyclosporine was injected once, on day 2 after transplantation. All heart allografts transplanted alone and treated with cyclosporine were rejected acutely (median survival time [MST] of 14.5 days). In contrast, after combined transplantation of a donor lung or spleen with the heart, almost all heart grafts survived indefinitely. Transplantation of a syngeneic lung or third-party spleen had little effect on heart graft survival (MST of 22.5 days and 26.5 days, respectively). Without cyclosporine treatment, combined transplantation with a donor lung or spleen hardly prolonged heart graft survival. Transplantation of a lymphocyte-depleted spleen with the heart induced a combi-effect in cyclosporine-treated rats that was somewhat weaker: only two of six hearts survived indefinitely. We conclude that in the combi-effect an immunological mechanism reduces rejection of the heart. This mechanism is probably generated by the lymphoid tissue (bronchus-associated lymphoid tissue in lung and white pulp in spleen) in the combined transplant.     

Evaluation of graft viability in heterotopic auxiliary liver transplantation in the rat.

In the rat model of heterotopic auxiliary liver transplantation, the coexistence of the engrafted liver and the recipient's native liver makes it difficult to evaluate the posttransplant graft viability. In this study, auxiliary liver transplantation was performed in Wistar rats, in which the recipient's native liver was handicapped with a 68% partial hepatectomy and a common bile duct ligation. Serum biochemistry of the liver was analyzed and compared with that of the selected control group. The surgical handicap of the liver showed severe damaging effects: the handicapped native livers appeared atrophic at autopsy, and no long-term animal survival could be achieved without an auxiliary liver transplantation. As the engrafted liver corrected the cholestasis of the handicapped native liver, significant differences of serum biochemistry were found between the transplanted group and the control group: for bilirubin concentration and gamma glutamyl transferase activity from postoperative day 3 to 28 (p < .05); for alkaline phosphatase on days 3, 7, 14, and 28 (p < .05); for alanine aminotransferase activity on days 3 and 14 (p < .05); and for aspartate aminotransferase activity on day 14 (p < .05). The efficiency to induce hepatic failure and to hamper its regeneration capacity in the native liver makes animal survival and liver biology as reliable parameters to evaluate the posttransplant graft viability in this rat model.     

大鼠颈部心脏移植中动脉套叠吻合的应用

目的探讨采用动脉套叠吻合结合套管连接技术,制作大鼠颈部心脏移植模型,以简化手术操作,提高成功率. 方法模型通过将供心无名动脉套叠吻合于受体右颈总动脉,供心肺动脉套接于受体右颈外静脉,将雄性SD大鼠(25只)心脏移植于Wistar大鼠(25只)颈部.术后应用环孢霉素A (1.5 mg/kg, q.d.)灌胃治疗,通过视诊和触诊观察移植心存活情况,以存活超过3天为手术成功标准,观察期3个月. 结果正式实验25例,平均手术时间为(48.7±3.4)分钟,手术成功22例,成功率为88%.失败3例为吻合口出血1例,血栓形成2例.观察期内死亡6例,原因为肺部感染、颈部脓肿、肝脏或膀胱肿瘤,其余16例大鼠移植心存活均超过3个月. 结论改进的大鼠颈部心脏移植手术模型具有操作简便,心脏缺血时间短,便于观察等优点.     

Auxiliary partial orthotopic living donor liver transplantation in a patient with alcoholic liver cirrhosis to overcome donor steatosis

The efficacy of auxiliary partial orthotopic liver transplantation (APOLT) to overcome the problems associated with a markedly steatotic graft in a living donor has not been fully explored. We have recently performed APOLT in a patient with alcoholic liver disease, where the only potential candidate donor was affected by 50% macrovesicular steatosis and 30% microvesicular steatosis. The recipient's left liver was resected and the donor's left liver, corresponding to a 0.46% graft-to-recipient weight ratio, was orthotopically transplanted. The postoperative course of this patient was uneventful, except for a transient large amount of ascites. Native liver volume in the recipient serially decreased, and the volume of the graft serially increased after transplantation. Four months after transplantation, the donor and recipient are doing well with a normal liver function. In conclusion, APOLT may be a feasible solution for a markedly steatotic living donor graft in patients with alcoholic liver disease.     

Break of tolerance via donor-specific blood transfusion by high doses of steroids: a differential effect after intestinal transplantation and heart transplantation

ABSTRACT: Tolerance requires active mechanisms. How immunosuppressors affects tolerance is poorly understood. METHODS: RA (RT1(p))/PVG (RT1(c)) rats were used as donor/recipient. Intestinal and heart transplant model were selected as highly and poorly immunogenic organs. Studied groups were 1, rejecting control; 2, received peritransplant steroids; 3, donor-specific blood transfusion (DSBT); 4, DSBT plus peritransplant steroids; and 5, DSBT+periDSBT Ste. RESULTS: Intestinal transplant recipients in group 1 died on posttransplant day (d) 18. In group 2, steroids did not change survival (17 days, P >.05 versus group 1). With DSBT (group 3), all rats survived >75 days, whereas with steroids those in group 4 survived 59 days (P >.05 vs group 3) and group 5 survived 51 days (P <.05 versus group 3). Survivors in group 2 were tolerant as evidenced by acceptance of secondary donor-specific (not third-party) graft. However, 100% and 33% of donor-specific secondary grafts were rejected in groups 4 and 5 (P <.05 and P >.05 versus group 3). In heart transplants, steroid treatment had no effect on graft survival (group 1 9 days; group 2 9 days; P >.05). DSBT (group 3) induced 100% tolerance (primary: >100 days, secondary: 100%). Unlike in intestinal transplantation, adjunction peritransplant steroids (group 4) allowed 100% of primary and 83% of secondary graft acceptance (P >.05 versus group 3). In group 5, (DSBT+periDSBT steroids) acceptance of primary and secondary grafts tended to be reduced (primary: 77 days; P >.05 versus group 3; secondary: 67%, P >.05 versus group 3). CONCLUSION: Steroid induction did not prolong graft survival after either intestinal or heart transplant. Adjunction of steroids to a DSBT tolerogenic regimen caused rejection of primary and secondary grafts, particularly after intestinal transplantation. Routine use of steroids in the clinics must be reconsidered, particularly when immunogenic organs are transplanted and when immunomodulation is applied.     

成人间活体扩大右半肝移植治疗急性肝功能衰竭

目的介绍成人间活体扩大右半肝移植治疗急性肝功能衰竭的临床经验。方法对1例42岁男性急性肝功能衰竭合并肝性脑病Ⅲ期患者行活体扩大右半肝移植治疗。其45岁姐姐为供者,CT评估供者包含肝中静脉的扩大右半肝体积为728．4cm^2（801g）,供肝／受者体重比为1．3％。供肝之肝右、中静脉整形后与受者整形后之肝右静脉行端-侧吻合;供受者门静脉、肝动脉行端．端吻合。供肝胆管整形后与受者胆总管行端-端吻合。结果供、受者手术均成功。供者术后恢复顺利,受者术后8h恢复意识,14d后丙氨酸转氨酶、总胆红素等指标首次下降至正常水平。术后16d曾出现转氨酶明显升高,给予甲泼尼龙1000mg冲击治疗后恢复正常。随访至今,供受者已健康生存8个月,均未出现胆管、肝动脉及静脉回流等并发症。结论扩大右半肝移植在技术上完全可行。能为成人患者提供足够重量的移植物,尤其对于急性肝功能衰竭患者具有重要意义,术前精确的影像学评估,熟练的肝切除和肝移植技术是确保该类手术成功的关键因素。     

Partial left ventricular unloading reverses contractile dysfunction and helps recover gene expressions in failing rat hearts

We investigated the effects of partial left ventricular unloading on failing rat hearts by using heterotopic heart-lung transplantation model. Heart failure (HF) was induced in Lewis rats by ligating the left anterior descending artery. After four weeks, the infarcted hearts and lungs were harvested and transplanted into the recipient rats by anastomosing donor's ascending aorta to recipient's abdominal aorta. Therefore, coronary venous blood entered the left ventricle (LV) and LV was partially unloaded (HF-PU group). Normal and infarcted heart rats (HF group) without transplantation served as control animals. After two weeks' unloading, the infarcted LV in HF-PU group significantly decreased its weight and myocardial diameter compared with HF group and they were close to normal levels. Developed tension of posterior papillary muscle was significantly increased in HF-PU group compared with HF group. The mRNA expressions of brain natriuretic peptide (BNP), sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a), beta(1) and beta(2)-adrenergic receptors (beta(1) and beta(2)-AR) in LV tissue were almost normalized in HF-PU group. Partial left ventricular unloading regressed myocardial hypertrophy, reversed contractile dysfunction and normalized the mRNA (BNP, SERCA2a, beta(1) and beta(2)-AR) expressions of failing rat hearts.     

大鼠小肠-辅助性肝脏联合移植的实验研究

目的　研究小肠—辅助性肝脏联合移植的可行性以及移植后的肝脏增生。方法　供、受者均为Lewis大鼠 ,分为移植组和对照组。移植组 :切取供者小肠和 6 0 %肝脏 ;切除受者小肠 ;然后将移植物植入受者。移植物血液供应通过连自移植物的腹主动脉和受者腹主动脉端 侧吻合 ,静脉回流通过移植物的肝下下腔静脉和受者左、右肾静脉之间的下腔静脉端 侧吻合。对照组 :单行剖腹探查 ;观察 1 8个月后处死。分别测量两组的肝脏重量 ,检测肝功能、肝脏和小肠的组织学改变。结果　移植组受者自身肝脏未见增生 ,移植肝脏明显增生 ;受者的肝脏 /体重比明显升高。移植组和对照组体重、肝脏功能检查、肝脏和小肠的组织学差异不显著。移植组受者自身肝脏、移植肝脏及对照组肝脏肝细胞的凋亡指数差异无显著性。移植组受者自身肝脏肝细胞增殖指数较移植肝脏及对照组肝脏肝细胞的增殖指数明显降低。结论　小肠—辅助性肝脏联合移植是可行的 ;辅助性肝移植可以提高肝脏 /体重比。     

Transfer of "infectious" cardiac allograft tolerance induced by donor-specific transfusion

BACKGROUND: "Infectious tolerance" has been defined as the tolerance induced in a new recipient by the adoptive transfer of cells from a recipient accepting an allograft after anti-CD4 and anti-CD8 monoclonal antibody treatment. A clear understanding of the mechanisms responsible for graft acceptance after donor-specific blood transfusion (DST) has remained elusive. We examined the development and "infectious" nature of immunologic changes resulting in indefinite survival of LEW to DA rat cardiac allografts after DST alone without the need for antibody. METHODS: One hundred x 10(6) LEW splenocytes (SC) as DST were injected intravenously into DA recipients 7 days before LEW cardiac transplantation. Subsequently, 100 x 10(6) SC harvested from a DA recipient 30, 60, or 100 days after graft acceptance were adoptively transferred into lightly gamma-irradiated (450 rad) na?ve DA recipients 24 hours before a second LEW cardiac allograft. Subsequent graft function was determined. RESULTS: Adoptive transfer of SC from the DST-treated DA rats 30 days after LEW heart transplant acceptance into na?ve gamma-irradiated DA rats failed to transfer tolerance to LEW cardiac allografts. However, SC from DA rats bearing LEW hearts for more than 60 days induced indefinite tolerance to all LEW hearts. This infectious tolerance could be adoptively transferred again to a second DA recipient. CONCLUSIONS: DST-generated regulatory cells can downregulate na?ve lymphocytes to promote allograft acceptance. This tolerance can be expanded and serially transferred to a subsequent na?ve cardiac recipient.     

脾切除对心脏移植大鼠外周血淋巴细胞凋亡及调节性T淋巴细胞的影响

目的 探讨脾切除对同种异体心脏移植大鼠外周血淋巴细胞凋亡及调节性T淋巴细胞的影响.方法 以Wistar大鼠为供者、SD大鼠为受者,进行腹部异位心脏移植,同时切除受者的脾脏(心脏移植切脾组),并以不切脾者为对照(心脏移植对照组),另设不行任何处理的对照组和单纯切脾的单纯切脾组.术后第1、3、5、7天.取各组受者的移植心脏和外周血,观察移植心脏的组织学变化和细胞超微结构改变情况,以流式细胞仪检测外周血淋巴细胞的凋亡率及CD4+ CD25+ T淋巴细胞的变化,逆转录聚合酶链反应检测CD4+ CD25+ T淋巴细胞上Foxp3 mRNA的表达情况,记录移植心脏的存活时间.结果 心脏移植对照组移植心脏存活时间为(7.47±2.24)d,心脏移植切脾组移植心脏存活时间为(17.63±4.54)d,二者间的差异有统计学意义(P<0.05).心脏移植对照组的移植心脏肿胀,质硬,色暗,间质水肿、出血,弥漫性炎症细胞浸润,大量心肌细胞坏死、溶解,横纹不清;心脏移植切脾组的移植心脏质软,色红,局部灰白,外膜下以及细胞间局灶性水肿,炎症细胞浸润,心肌细胞结构完整,横纹清晰;心脏移植切脾组的细胞超微结构改变轻于心脏移植对照组.心脏移植切脾组术后第5天和第7天的淋巴细胞凋亡率分别为(7.62±2.15)%和(9.41±3.82)%,明显高于心脏移植对照组(P<0.05,P<0.05).心脏移植切脾组术后第3、5、7天时的CD4+ CD25+ T淋巴细胞明显多于心脏移植对照组(P<0.01,P<0.01,P<0.01),其Foxp3 mRNA的表达也较心脏移植对照组明显上调.结论 脾切除使心脏移植大鼠外周血淋巴细胞凋亡率增加,调节性T淋巴细胞增多,其Foxp3 mRNA表达上调,这些变化与移植心脏病理改变呈负相关.     

Auxiliary partial liver grafting in rats: effect of host hepatectomy on graft regeneration,and review of literature on surgical technique

Auxiliary partial liver transplantation (APLT) is beneficial for fulminant liver failure when there is potential for recovery of the diseased liver. However, the impact of host hepatectomy on regeneration of the grafted liver is unclear. In this study, we modified a previous rat model of auxiliary whole liver transplantation without portal vein reconstruction, and studied the effect of host hepatectomy on regeneration of the cut liver graft. Thirty percent of the liver was heterotopically transplanted, to connect the recipient's left renal artery and vein with the graft's aortic cuff of the hepatic artery and inferior vena cava, respectively, using a cuff technique; 30% of the recipient liver then was cut. The control group was left intact. The liver grafts were weighed preoperatively and 2 weeks postoperatively. This procedure prevented congestion of the graft liver and achieved a high success rate, even when performed by a surgeon who was relatively inexperienced with the technique. The weight of the grafted liver in the host hepatectomized group significantly increased (P < 0.05) compared with that of the control group. We developed an experimental model of APLT and reviewed the literature on rat heterotopic liver transplantation, and compared the surgical techniques.     

Long-term survival of xenogeneic heart grafts achieved by costimulatory blockade and transient mixed chimerism

BACKGROUND: Xenotransplantation holds great promise in clinical medicine, but is limited by the vigorous rejection response elicited against solid organs transplanted across species barriers. In this study, we investigated the role of anti-CD40L monoclonal antibody (mAb) in inducing xenogeneic mixed chimerism and donor-specific heart transplantation tolerance. METHODS: One day before heart transplantation, mice were injected intraperitoneally with anti-mouse CD8/NK1.1/Thy1.2 mAbs. On day 0, the mice received 3 Gy total body irradiation (TBI), an intravenous injection of unseparated bone marrow (BM) harvested from F344 rats, and an intraperitoneal injection of hamster antimouse CD40L mAb, MR1. Heart grafts from F344 rats were heterotopically transplanted into the abdomen of B6 mouse recipients. Using flow cytometric analysis of peripheral white blood cells, we assessed donor hematopoiesis at various times after bone marrow transplantation (BMT). RESULTS: Chimerism subsided gradually and disappeared completely 18 weeks after BMT. The cardiac graft survived permanently, even after the mixed chimerism disappeared. To determine if the mice acquired donor-specific tolerance, second rat heart grafts were transplanted 120 days after the first heart transplantation. The second transplanted hearts were also accepted over 60 days. Histological analysis revealed no remarkable vasculopathy in the coronary vessels at any stage. CONCLUSIONS: These findings clearly show that costimulatory blockade plays an important role in inducing xenochimerism, and that transient mixed chimerism can induce permanent acceptance of rat to mouse cardiac xenografts. Transplantation of xenogeneic bone marrow cells under costimulatory blockade at the time of heart transplantation may induce transplantation tolerance.     

Repopulation of donor heart by recipient bone marrow-derived dendritic cells prior to transplantation causes acute rejection by both the allogeneic and syngeneic recipient

Experimental studies on allogeneic transplantation have shown that recipient dendritic cells (DC) play a role in peripheral tolerance as well as in rejection of allografts. It is not known whether DC exert their tolerogenic function in recipient lymphoid tissue, and whether they process shed alloantigen in the graft itself. To answer this question we created a chimeric heart model deprived of its own DC and repopulated by recipient DC. The rationale for this model was to observe whether recipient DC located in the graft attenuate recruitment and stimulation of recipient lymphocytes, subsequently prolonging graft survival. Vascularized bone marrow transplants (VBMTx) from the prospective recipient to the lethally irradiated heart donor, which function for a period of 14 days, were used to replace donor DC with prospective recipient DC. Hearts from chimeric LEW rats (with BN DC) were transplanted to untreated BN rats. Also, hearts from chimeric LEW rats (with BN DC) were returned to untreated LEW rats. Replacement of the donor heart with recipient DC did not prolong graft survival. Rather, it initiated a rejection reaction that was already present in the donor. Recipient DC retained their immunogenic properties also when the graft was returned back to a donor strain animal.     

A novel auxiliary partial orthotopic liver transplantation model in rats

BACKGROUND: Although auxiliary partial orthotopic liver transplantation (APOLT) has become a well-accepted procedure recently, a practical experiment model in APOLT using small animals has yet to be developed. METHODS: Male Lewis rats were used for both donors and recipients. An auxiliary partial graft was obtained by ex vivo resection of the donor right and caudate lobes, and was transplanted orthotopically into the recipient after resection of the recipient medial and left hepatic lobes. Portal vein and hepatic duct reconstructions were by the cuff technique, and supra- and intrahepatic vena cava were sutured continuously. Operative outcomes, serum chemistry, liver tissue blood flow, angiographic and histopathological findings were then examined. Conventional orthotopic liver transplantation (OLT) procedures were also undertaken as a control. RESULTS: One-day, 1-week and 1-month survival rate of APOLT group was 100, 85 and 85%, respectively. AST in the APOLT group on the 1st postoperative day was significantly higher than in the OLT group. No significant differences were recognized in serum albumin and total bilirubin levels between the two groups. Although the portogram of an APOLT rat showed slight narrowing at the cuff anastomosis site, both the graft and the native liver were opacified similarly. The liver tissue blood flow on the 5th postoperative day in the native liver and the graft returned to as high as 95 and 74% of the values on laparotomy, respectively. Histological examinations of the auxiliary graft 1 month after transplantation showed mild ductular proliferation and mononuclear cell infiltration around the portal triads. CONCLUSION: This novel APOLT model in rats allows practical and reproducible results, and may be of value in the basic study of APOLT procedures.     

Abdominal heart transplantation: description of a new allograft extrathoracic pump

This study describes a new kind of abdominal heart transplantation for left ventricular assistance carried out in a series of 12 experiments in pigs weighing 15-25 kg. This was achieved making three connections between the donor's left atrium, aorta and pulmonary artery with the recipient's aorta, still aorta and inferior vena cava, respectively. The hemodynamic data were satisfactory, the best survival rate with a transplanted working heart was 1 month. The low output of the recipient's left ventricle was obtained by ligation of the left anterior descending (LAD) coronary artery. In all animals, the highest peak of the pressure of the transplanted left ventricle was at least 20 mm Hg higher (ranging from 20 to 60 mm Hg) than the pressure in the recipient's left ventricle, and corresponded with the peak of the systemic arterial pressure. The cardiac output of the transplanted hearts showed a good hemodynamic response with support of the circulation after ligation of the LAD coronary artery in the recipient's heart.     

Auxiliary liver transplantation in acute liver failure in the rat – an illustrated description of a new surgical approach

INTRODUCTION: To investigate auxiliary liver transplantation successfully in rats suffering from acute liver failure, we developed a new surgical approach. METHODS: A 70% hepatectomized liver graft was implanted into the right upper quadrant of the abdomen. The donor portal vein was anastomosed with the recipient's right renal artery using the splint technique. The donor infrahepatic vena cava was attached onto the recipient vena cava end to side. The bile duct was implanted into the duodenum.     

Technique for retransplanting heterotopic heart grafts in mice

Removal of a transplanted organ from its original recipient and retransplanting it into a new host is an important method to study the role of the graft in the rejection process. Here we describe a novel technique of heart retransplantation in the mouse. In this technique, a primarily vascularized heart graft is anastomosed to the abdominal aorta and inferior vena cava of a syngeneic or immunodeficient allogeneic mouse, using standard techniques. Either 10 or 70 days later, the same graft is retransplanted into the abdomen of a second mouse by end-to-side anastomosis of the donor (first recipient) aortic and inferior vena cava's cuffs to the second recipient's abdominal aorta and inferior vena cava, respectively. A greater than 90% success rate was achieved by using this microsurgical technique. This method should be useful for studying intragraft factors, such as ischemia-reperfusion injury and donor antigen-presenting cells, on the outcomes of transplantations.     

Reloading the heart: a new animal model of left ventricular assist device removal

OBJECTIVES: Left ventricular assist devices are proposed as a bridge to recovery, but recurrent ventricular deterioration has limited this approach. We describe a new animal model that simulates the effects of left ventricular assist device unloading and then reloading after device removal. The model might facilitate the evaluation of interventions intended to prevent recurrent ventricular dysfunction. METHODS: The hearts and lungs of Lewis rats were removed and transplanted into the abdomen of recipient rats by anastomosing the donor's ascending aorta to the recipient's abdominal aorta. The transplanted hearts were maintained unloaded for 2 weeks in 49 animals. Eighteen transplanted hearts were removed after 2 weeks of unloading. In 17 animals the donor's right pulmonary artery was anastomosed to the recipient's abdominal aorta to reload the heart for an additional 2 weeks. In 14 animals the hearts were maintained unloaded for 4 weeks (an additional 2 weeks). The unloaded and reloaded hearts were compared with normal rat hearts (n = 18). RESULTS: In the unloaded hearts the left ventricular end-diastolic pressures remained low. The left ventricular systolic pressures were lower than the aortic pressures. The left ventricular weights (n = 8) and volumes (n = 4) remained significantly lower ( P < .01) than in the normal hearts. Two weeks after reloading, the left ventricular end-diastolic pressure (n = 8) increased ( P < .01), and the ventricle ejected. The left ventricular systolic pressures exceeded the aortic pressures. The left ventricular weights and volumes increased ( P < .01) and approached those of normal hearts. Matrix metalloproteinase 9 (n = 6/group) levels decreased in the unloaded state ( P = .02) and increased back to normal values after reloading. CONCLUSIONS: This surgical model simulated left ventricular assist device unloading of the left ventricle. The second operation reloaded the left ventricle, which then enlarged. This model will permit the evaluation of adjunctive interventions, such as cell transplantation, intended to facilitate successful left ventricular assist device removal and prevent recurrent dilatation.     

A simple new model of physiologically working heterotopic rat heart transplantation provides hemodynamic performance equivalent to that of an orthotopic heart.

BACKGROUND: The widely used non-volume-loaded abdominal heterotopic heart transplant (NL) in rats undergoes atrophy after transplantation. Various techniques have been designed to load the transplanted heart because of its potential immunological impact. Our aim was to create a volume-loaded heterotopic heart transplantation model (VL) capable of ejection and practical for routine studies. Using this model, we tested the hypothesis that VL isografts would retain myocardial performance comparable to native hearts (NH). METHODS: Heterotopic hearts were transplanted using and end-to-side anastomosis between the donor's superior vena cava and the recipient's abdominal inferior vena cava. The right ventricle loads the left ventricle (LV) via a direct anastomosis of the pulmonary artery to the left atrium. The LV ejects volume through an end-to-side anastomosis of the donor's aorta to the recipient's abdominal aorta. Hemodynamic data (systolic and diastolic LV pressures, dP/dt max and min, tau) were studied in-situ (at baseline and after adding volume) and in a Langendorff perfusion system (at baseline and after stimulation with isoproterenol) 2 weeks after transplantation.RESULTS: In situ systolic pressure and diastolic function of VL was superior to NL, and beta-adrenergic stimulated performance in the Langendorff perfusion of VL showed hemodynamic performance equivalent to NH, unlike NL which had a diminished response. CONCLUSION: This technique results in a volume-loaded ejecting heart transplant model that preserves anatomical structures. The VL can be evaluated in situ and after explantation in Langendorff perfusion system and may offer advantages if workload of the graft is of significance to the study performed.