An improved animal model of orthotopic liver transplantation in swine

Objective: To establish a swine model of orthotopic liver transplantation (OLT) which has high standardization, superior reproducibility and stability. Methods: The rate of success, reproducibility and stability were investigated on the modification of OLTs in closed miniature swine with series of improvements. Results: 20 OLTs were performed on the basis of improvements in experimental animals, surgical procedures and operative monitorings. The mean operation time and anhepatic phase was (181±25.8) and (28.4±3.2) min respectively, which were significantly shorter than those of the previous reports. Liver function of the animals recovered shortly after operation. One-week survival rate was 90%, and 15 animals survived more than 1 month. The incidence of vascular and biliary complications was lower in animals with long-term survival. Conclusion: The improved animal model of OLTs in swine is easy to operate with high standardization and rate of success, superior reproducibility and stability. It is an ideal model for series studies related to liver transplantation in big animals.

Using Tree Shrews (Tupaia belangeri)as a Novel Animal Model of Liver Transplantation

Liver transplantation (LT)is most effective and promising approach for end-stage liver disease.However,there remains room for further improvement and innovation,for example, to reduce ischemic reperfusion injury,transplant rejection and immune tolerance.A good animal model of LT is essential for such innovation in transplant research.Although rat LT model has been used since the last century,it has never been an ideal model because the results observed in rat may not be applied to human because these two species are genetically distinct from each other.In this study,we for the first time performed LT using the tree shrew (Tupaia belangeri),a species in the Order Scandentia which is closely related with primates,and evaluated the possibility to adopt this species as a new model of LT.We performed LT on 30 animals using the two-cuff technique, examining the success rate,the survival rate and the immunological reaction.The recipient operation time was 60 min averagely,and we limited the time of the anhepatic phase within 20 min.Twenty-seven (90%)of the animals survived for at least 3 days after the transplantation. Thirteen animals that did not receive any immunosuppressive drug died in 8 days mostly because of acute rejection effect (n=9),similar to the reaction in human but not in experimental rat.The rest 14 animals that were given rapamycin survived significantly longer (38 days)and half of them survived for 60 days until the end of the study.Our results suggest that performing LT in tree shrews can yield high success rate and high survival rate.More importantly,the tree shrews share similar immunological reaction with human.In addition,previous genomics study found that the tree shrews share more proteins with human.In sum,the tree shrews may outperform the experimental rats and could be used as a better and cost-effective animal model for LT.

Modified arterialization of orthotopic liver transplantation in a mouse model

BACKGROUND: With the establishment of genetically modified and gene knock-out models, the mouse has become an important animal model for liver transplantation. We examined hepatic rearterialization after liver transplantation in a mouse model. METHODS: Orthotopic liver transplantation was performed in 70 mice and sham-operation was performed in a control group of 40 mice. Based on the 'two-cuff' method, a continuous suture approach was applied to the suprahepatic inferior vena cava and a cuff approach to the portal vein and the infrahepatic inferior vena cava. A biliary stent was inserted into the bile duct. The hepatic artery was reconstructed with end-to-side anastomosis. The survival rate of recipients was monitored at 24 hours, one week, and one month after the operation. Liver function and morphology were evaluated one month postoperatively. RESULTS: Postoperative survival rates were 94.3% at 24 hours, 91.4% at one week, and 85.7% at one month. No significant difference was seen between the experimental and control groups in liver function. The hepatic tissue preserved normal structure. CONCLUSION: Owing to its high survival rate and stability, this surgical approach is ideal for establishing an orthotopic liver transplantation mouse model with hepatic artery reconstruction. (Hepatobiliary Pancreat Dis Jut 2010; 9: 264-268)

Establishment of a new pig model for auxiliary partial orthotopic liver transplantation

AIM: To establish a new pig model for auxiliary partial orthotopic liver transplantation (APOLT).METHODS: The liver of the donor was removed from its body. The left lobe of the liver was resected in vivo and the right lobe was used as a graft. After the left lateral lobe of the recipient was resected, end-to-side anastomoses of suprahepatic inferior vena cava and portal vein were performed between the donor and recipient livers,respectively. End-to-end anastomoses were made between hepatic artery of graft and splenic artery of the host.Outside drainage was placed in donor common bile duct.RESULTS: Models of APOLT were established in 5 pigs with a success rate of 80%. Color ultrasound examination showed an increase of blood flow of graft on 5th d compared to the first day after operation. When animals were killed on the 5th d after operation, thrombosis of hepatic vein (HV) and portal vein (PV) were not found. Histopathological examination of liver samples revealed evidence of damage with mild steatosis and sporadic necrotic hepatocytes and focal hepatic lobules structure disorganized in graft. Infiltration of inflammatory cells was mild in portal or central vein area. Hematologic laboratory values and blood chemical findings revealed that compared with group A (before transplantation), mean arterial pressure (MAP), central venous pressure (CVP), buffer base (BB), standard bicarbonate (SB) and K+ in group B (after portal vein was clamped) decreased (P＜0.01). After reperfusion of the graft, MAP, CVP and K+ restored gradually.CONCLUSION: Significant decrease of congestion in portal vein and shortened blocking time were obtained because of the application of in vitro veno-venous bypass during complete vascular clamping. This new procedure,with such advantages as simple vessel processing, quality anastomosis, less postoperative hemorrhage and higher success rate, effectively prevents ischemia reperfusion injury of the host liver and deserves to be spread.

Details determining the success in establishing a mouse orthotopic liver transplantation model

Liver transplantation(LT)is currently the only effective treatment option for endstage liver disease.The importance of animal models in transplantation is widely recognized among researchers.Because of the well-characterized mouse genome and the greater diversity and availability of both genetically modified animals and research reagents,mouse orthotopic LT(MOLT)has become an ideal model for the investigation of liver biology,tissue injury,regulation of alloimmunity and tolerance induction,and the pathogenesis of specific liver diseases.However,due to its complicated and technically demanding procedure,the model has merely been used by only a few research groups in the world for years.For a new learner,training lasting at least a couple of months or even years is required.Most of the investigators have emphasized the importance of elaborate techniques and dedicated instruments in establishing a MOLT model,but some details are often neglected.The nontechnical details are also significant,especially for researchers who have little experience in mouse microsurgery.Here,we review and summarize the crucial technical and nontechnical details in establishing the model of MOLT based on scientific articles and our experience in six aspects:animal selection,anesthesia,perioperative management,organ procurement,back-table preparation,and implantation surgery.We aim to enable research groups to shorten the learning curve and implement the mouse LT procedure with high technical success.

Improved two-cuff technique for orthotopic liver transplantation in rat

BACKGROUND: The first orthotopic liver transplantatio in rat (ROLT) was reported by Lee in 1973. Kamada inno vatively applied cuff technique to ROLT in 1979. However the operative procedures were highly demanding and th operative mortality was relatively high. The purpose of thi study was to improve the model of ROLT, simplify opera tive procedures, and enhance the successful rate of opera tion. METHODS: Orthotopic liver transplantation was per formed in 160 Wistar rats by improved two-cuff technique The portal vein between donor and recipient was anasto mosed with the cuff technique. The same method was use to anastomose the infrahepatic vena cava. The suprahepati vena cava and the hepatic artery were anastomosed by m crovascular suturing and the bile duct was anastomosed en to end by a Teflon catheter. RESULTS: The average time for donor operation, graf preparation and anhepatic phase was 31 minutes, 14 mi nutes and 13 minutes, respectively. The anastomosis tim for the suprahepatic vena cava, portal vein, infrahepatic ve na cava, hepatic artery and bile duct was 7 minutes, 2 mi nutes, 2 minutes, 8 minutes and 1 minute, respectively The main causes for operative mortality were pneumotho rax, anesthesia, air embolism and massive bleeding, an the successful rate of operation was 92.5%. The causes fo death after operation were stoma bleeding, infection, b liary obstruction and graft failure. CONCLUSION: The improved two-cuff technique can re duce operative mortality, enhance survival rate, and serv as an ideal method for the establishment of animal model o ROLT.

Orthotopic liver transplantation for giant liver haemangioma: A case report

In liver haemangiomas, the risk of complication rises with increasing size, and treatment can be obligatory. Here we present a case of a 46-year-old female who suffered from a giant haemangioma causing severe portal hypertension and vena cava compression, leading to therapy refractory ascites, hyponatremia and venostasis-associated thrombosis with pulmonary embolism. The patients did not experience tumour rupture or consumptive coagulopathy. Surgical resection was impossible because of steatosis of the non-affected liver. Orthotopic liver transplantation was identified as the only treatment option. The patient's renal function remained stable even though progressive morbidity and organ allocation were improbable according to the patient's lab model for end-stage liver disease(lab MELD) score. Therefore, non-standard exception status was approved by the European organ allocation network "Eurotransplant". The patient underwent successful orthotopic liver transplantation 16 mo after admission to our centre. Our case report indicates the underrepresentation of morbidity associated with refractory ascites in the lab MELD-based transplant allocation system, and it indicates the necessity of promptly applying for non-standard exception status to enable transplantation in patients with a severe clinical condition but low lab MELD score. Our case highlights the fact that liver transplantation should be considered early in patients with non-resectable, symptomatic benign liver tumours.

Biliary tract injury caused by different relative warm ischemia time in liver transplantation in rats

BACKGROUND: There is a controversy over the degree of liver and biliary injury caused by the period of secondary warm ischemia. A liver autotransplantation model was adopted because it excludes the effects of infection and immunological rejection on bile duct injury. This study was undertaken to assess biliary tract injury caused by relative warm ischemia (secondary warm ischemia time in the biliary tract) and reperfusion. METHODS: One hundred and two rats were randomly divided into 5 groups: group I (control); groups 11 to V, relative warm ischemia times of 0 minute, 30 minutes, I hour and 2 hours. In addition to the levels of serum alkaline phosphatase, and total bilirubin, pathomorphology assessment and TUNEL assay were performed to evaluate biliary tract damage. RESULTS: Under the conditions that there were no significant differences in warm ischemia time, cold perfusion time and anhepatic phase, group comparisons showed statistically significant differences. The least injury occurred in group H (portal vein and hepatic artery reperfused simultaneously) but the most severe injury occurred in group V (biliary tract relative warm ischemia time 2 hours). CONCLUSIONS: Relative warm ischemia is one of the factors that result in bile duct injury, and the relationship between relative warm ischemia time the bile injury degree is time-dependent. Simultaneous arterial and portal reperfusion is the best choice to avoid the bile duct injury caused by relative warm ischemia. (Hepatobiliary Pancreat Dis Int 2009; 8: 247-254)

Comprehensive and innovative techniques for livertransplantation in rats: A surgical guide

AIM: To investigate our learning curves of orthotopic liver transplantation (OLT) in rats and the most important factor for successful surgery. METHODS: We describe the surgical procedures for our rat OLT model, and determined the operator learning curves. The various factors that contributed to successful surgery were determined. The most important surgical factors were evaluated between successful and unsuccessful surgeries.RESULTS: Learning curve data indicated that 50 cases were required for operator training to start a study. Operative time, blood loss, warm ischemic time, anhepatic phase, unstable systemic hemodynamic state, and body temperature after surgery significantly affected surgery success by univariate analysis, while the anhepatic phase was the most critical factor for success by multivariate analysis. CONCLUSION: OLT in rats is the only liver transplantation model that provides clinically relevant and reliable results. Shortened anhepatic phase is key to success in this model.

Liver cell transplantation for Crigler-Najjar syndrome type Ⅰ: Update and perspectives

Liver cell transplantation is an attractive technique to treat liver-based inborn errors of metabolism. The feasibility and efficacy of the procedure has been demonstrated, leading to medium term partial metabolic control of various diseases. Crigler-Najjar is the paradigm of such diseases in that the host liver is lacking one function with an otherwise normal parenchyma. The patient is at permanent risk for irreversible brain damage. The goal of liver cell transplantation is to reduce serum bilirubin levels within safe limits and to alleviate phototherapy requirements to improve quality of life. Preliminary data on Gunn rats, the rodent model of the disease, were encouraging and have led to successful clinical trials. Herein we report on two additional patients and describe the current limits of the technique in terms of durability of the response as compared to alternative therapeutic procedures. We discuss the future developments of the technique and new emerging perspectives.

Establishment of an animal model of ischemic type intrahepatic biliary lesion in rabbits

AIM:To explore a method to establish an animal model of ischemic type intrahepatic biliary lesion in rabbits. METHODS:Forty Japanese white rabbits of clean grade were divided randomly into four groups(10 rabbits per group)including sham operation(SO) group,and artery-bile obstruction(ABO)-1 h group, ABO-2 h group and ABO-3 h group.All the rabbits in this study underwent the same initial surgical procedure in which the liver was prepared as for graft removal during liver transplantation.Subsequently in the SO group,no additional vascular intervention was performed,while in groups ABO-1 h,ABO-2 h and ABO-3 h,the animals underwent combined clamping of the hepatic artery and common bile duct with microvascular clips for 1,2 and 3 h,respectively.After the scheduled occlusion time,the clip was removed to recover blood supply.The animals were killed 4 wk after operation.The survival rate,liver function, cholangiography and histopathological manifestation of the rabbits in each group were observed. RESULTS:The survival rate was 100%in groups SO,ABO-1 h and ABO-2 h,while it was 60%in group ABO-3 h.At each observation time,the change degree of the indexes of liver function was proportional to the clamping time(ABO-3 h>ABO-2 h>ABO-1 h> SO,P<0.05).Cholangiographical and histopathologic manifestations both showed that intrahepatic biliary lesion aggravated proportionally with the increase of the clamping time. CONCLUSION:An animal model of ischemic type intrahepatic biliary lesion in rabbits is successfully established,which may provide a reliable technique for basic and clinical research into the etiology, development and prophylaxis of ischemic type intrahepatic biliary lesion after liver transplantation.

High incidence of biliary complications in rat liver transplantation:Can we avoid it?

MM： To investigate how to reduce the incidence of biliary complications in rat orthotopic liver transplantation. METHODS： A total of 165 male Wistar rats were ran- domly divided into three groups： Group A, orthotropic liver transplantation with modified ＂two-cuff＂ technique; Group B, bile duct was cut and reconstructed without transplantation; and Group C, only laparotomy was performed. Based on the approaches used for biliary reconstruction, Group A was divided into two sub-groups： A1 （n = 30）, duct-duct reconstruction, and A2 （n = 30）, duct-duodenum reconstruction. To study the influence of artery reconstruction on bile duct complication, Group B was divided into four sub-groups： B1 （n = 10）, duct-duct reconstruction with hepatic artery ligation, B2 （n = 10）, duct-duct reconstruction without hepatic artery ligation, B3 （n = 10）, duct-duodenum reconstruction with hepatic artery ligation, and B4 （n = 10）, duct-duodenum recon- struction without hepatic artery ligation. The samples were harvested 14 d after operation or at the time when significant biliary complication was found. RESULTS： In Group A, the anhepatic phase was 13.7 ＋ 1.06 min, and cold ischemia time was 50.5 ＋ 8.6 min. There was no significant difference between A1 and A2 in the operation duration. The time for biliary reconstruction was almost the same among all groups. The success rate for transplantation was 98.3% （59/60）. Significant differ- ences were found in the incidence of biliary complications in Groups A （41.7%）, B （27.5%） and C （0%）. A2 was more likely to have biliary complications than A1 （50% vs 33.3%）. B3 had the highest incidence of biliary complica- tions in Group B. CONCLUSION： Biliary complications are almost in- evitable using the classical ＂two cuff＂ techniques, and duct-duodenum reconstruction is not an ideal option in rat orthotopic liver transplantation.

BUILDING OF A SIMPLE MODEL OF AUTO-OLT IN DOGS AND ITS SIGNIFICANCE

AIM: we recommend a method of Simple auto-oLT model in dogs.METHODS:The model was ligated all ligaments or connective tissues of the liver,only reserved the vascular construction,that was the suprahepatic and infrahepatic inferior vena cava,portal vein,hepatic artery or common bile duct.the operation was similar to the orthotopic liver transplantation except vascular anastomoses,the dog liver underwent the warm or cold ischemia and the reperfusated injurous process.RESULTS: The imitability was exactly good and the operation was simple and safe. Because the hepatic vessels of the going out or coming in was clamped block and might open or blind the blood flow whenever necessary,the model might control the warm or cold ischemic time accurately,and eliminate the influence or the complications due to vascular anastomoses.CONCLUSIONS: The model avoided many-sided Influences of the traditional OLT and was a good method to study hepatic artery or portal vein ischemic injury and created a new way to explore the pathogenesis or some complications in the OLT.

3种方法构建BALB/c裸鼠BEL-7402人肝癌模型

【目的】以3种不同方法构建BALB/c裸鼠BEL-7402人肝癌模型,并观察BALB/c裸鼠肿瘤生长情况。【方法】将51只BALB/c裸鼠分为A、B、C 3组,每组17只。A组动物皮下注射0.2 m L BEL-7402人肝癌细胞,B组动物于肝脏原位注射0.05 mL BEL-7402人肝癌细胞与Matrigel Matrix的混合物,C组动物于肝脏原位接种BEL-7402人肝癌细胞瘤块组织。从接种后第2周至第5周,每周测定裸鼠体质量,且每组随机取3只动物解剖取肿瘤测量其大小和质量。第5周后,观察记录各组剩余动物(每组5只)的一般状态和存活天数。采用苏木素—伊红(HE)染色观察肿瘤组织病理学变化。【结果】A、B、C 3组实验动物成瘤率均为100%。5周后,A、B、C 3组剩余动物存活天数分别为(90.8±10.2)、(75.6±14.0)、(67.4±11.1) d。解剖所取肿瘤呈结节状,组织病理学检查可见肿瘤细胞呈一定方向排列,细胞呈梭形、多边形,异型明显,可见核分裂。【结论】本实验所采用的3种不同方法均能成功建立BALB/c裸鼠BEL-7402人肝癌模型。

HepaRG细胞移植治疗鼠特异性Fas抗体诱导的小鼠急性肝衰竭

目的探讨HepaRG细胞移植对鼠特异性Fas抗体诱导的急性肝衰竭小鼠的治疗作用和人肝细胞异种移植的策略。方法利用特异性鼠抗Fas抗体(Jo2mAb)腹腔注射,剂量为0.3mg/kg,诱导20只SCID小鼠制备急性肝衰竭模型,24h内按照实验动物随机数字表分组方法,经脾移植2×106HepaRG细胞的小鼠为实验组(10只),未经HepaRG细胞移植的小鼠为对照组(10只)。观察小鼠的存活率、肝脏功能、肝组织变化情况。实验组移植4周免疫组化检测肝组织人白蛋白、人CK18、人HepPar1的表达,免疫荧光检测人白蛋白的表达。结果实验组小鼠有9只存活超过4周,而对照组小鼠3d内先后死亡9只,实验组血清ALT和AST趋于正常,显著低于对照组(P<0.01),且存活时间显著高于对照组(P<0.01)。实验组经HepaRG细胞移植能避免Jo2mAb所致肝组织出血、坏死,移植后4周免疫组化可见人白蛋白、CK18、HepPar1阳性表达细胞,肝组织免疫荧光可见人白蛋白阳性细胞的表达。结论移植HepaRG细胞可治疗Jo2mAb腹腔注射小鼠诱导的急性肝衰竭,HepaRG细胞不仅在小鼠肝内存活,且得以增殖。

胆管上皮细胞分泌的VEGF-A在热缺血再灌注损伤后胆道及其微循环再生与修复中的作用

目的探讨热缺血再灌注损伤后血管内皮生长因子-A(vascular endothelial growth factor-A,VEGF-A)在胆道及其微循环再生与修复中的作用,为积极预防和治疗胆道热缺血再灌注损伤导致的胆道并发症提供理论依据。方法供受体均为雄性SD大鼠。供肝分别经历0min(对照组)和10min(实验组)热缺血后,进行重建肝动脉血供的大鼠肝移植模型。分别于供肝冷保存后(即0h)以及术后6h、24h、3d、7d、14d、30d处死大鼠,收集标本。测定血清ALT、GGT、TBIL。免疫组化法检测VEGF-A表达情况。分别标记胆管上皮细胞、血管内皮细胞、增殖细胞,统计汇管区胆管数、血管数以及胆管上皮细胞增殖指数。原位杂交法检测VEGF-A mRNA表达情况。结果实验组GGT和TBIL较对照组升高持续时间更长,术后30d和14d才降至正常。实验组汇管区胆管数及血管数均在术后14d达到最高值,且明显多于对照组。肝细胞和血管内皮细胞VEGF-A、VEGF-AmRNA表达在术后24h达到峰值后迅速下降,而胆管上皮细胞VEGF-A、VEGF-A mRNA表达持续升高,直到术后7d才达到峰值,并且实验组表达水平均强于相应时间点对照组的表达。结论热缺血再灌注损伤后,胆管上皮细胞合成和分泌VEGF-A增加,并且可能在促进胆管及胆管周围血管丛的再生中发挥重要作用。

丹参酮Ⅱ_A磺酸钠联合思美泰对同种异体1/2原位肝脏移植大鼠P450及肝功能的影响

目的采用改良双袖套法建立大鼠同种异体1/2原位肝脏移植动物模型,观察术后丹参酮Ⅱ_A磺酸钠对细胞色素P450亚型酶活性及肝功能的影响,探讨药物干预机制,为临床治疗肝移植合理用药提供理论依据。方法选择60只大鼠,随机分为空白对照组、模型组、思美泰组及丹参酮组,思美泰组经尾静脉注射思美泰2 mg/(kg·d),丹参酮组在此基础上经尾静脉注射丹参酮Ⅱ_A磺酸钠5 mg/(kg·d)治疗14 d,空白对照组及模型组注射等体积生理盐水。采用高效液相色谱仪检测治疗前后肝微粒体细胞色素P450亚型酶(CYP2E1、CYP1A2、CYP3A4、CYP2D6、CYP2C9),生化检测肝功能中胆汁酸(TBA)、总胆红素(STB)、天冬氨酸氨基转移酶(AST)及丙氨酸氨基转移酶(ALT)、谷氨酰转移酶(GGT)水平,并计算肝移植术大鼠存活率,评价丹参酮Ⅱ_A磺酸钠对大鼠同种异体1/2原位肝脏移植的影响。结果经丹参酮Ⅱ_A磺酸钠及思美泰药物干预,丹参酮组CYP2E1、CYP1A2、CYP2C9、CYP3A4酶活性及TBA、GGT、ALT、AST、STB均明显降低,存活率达60%,与模型组及思美泰组比较,差异均有统计学意义(P<0.05)。结论丹参酮Ⅱ_A磺酸钠抑制大鼠肝微粒体细胞色素P450亚型酶活性,减轻过敏反应,提高同种异体1/2原位肝脏移植大鼠存活率,恢复肝功能,与思美泰联用可增强Na+/K+-ATP酶泵功能,同时,活血化淤、增加肝脏组织供氧相关。

利用bcl-2抗Fas凋亡特性联合Fas抗体促进人鼠嵌合肝中人肝细胞的增殖

目的移植bcl-2转基因HepaRG细胞到小鼠体内,并联合使用Jo2抗体诱导鼠肝细胞凋亡策略,促进人鼠嵌合肝中人肝细胞的增殖。方法SCID鼠经脾移植2×106bcl-2转基因HepaRG细胞,移植后1d始腹腔注射0.2mg/kgJo2抗体,每周1次,持续10周为实验组;移植未转染bcl-2基因的HepaRG细胞,同样给予Jo2抗体腹腔注射为对照1组,移植未转染bcl-2基因的HepaRG细胞,未腹腔注射Jo2抗体为对照2组,建立人鼠嵌合肝动物模型。从2周开始,后4、8、12、16、20、24周,分别采用免疫组化、免疫荧光和逆转录聚合酶链反应(RT-PCR)检测不同时相点实验组和对照组嵌合肝中肝组织人清蛋白以及人清蛋白mRNA,并用酶联免疫法(ELISA)定量检测鼠血清人清蛋白的含量。结果实验组和对照组小鼠均能存活至24周,鼠肝组织和血清中均能检测到清蛋白的表达。实验组肝组织和血清人清蛋白的表达均可持续到24周,人血清清蛋白高峰值为95.32ng/mL,出现在16周;而对照1组肝组织和血清人清蛋白的表达可持续到20周,人血清清蛋白高峰值为42.37ng/mL,出现在12周;对照2组肝组织和血清人清蛋白的表达可持续到12周,人血清清蛋白高峰值为22.91ng/mL,出现在8周。结论利用bcl-2抗Fas凋亡特性,转染bcl-2基因的HepaRG细胞移植并联合小剂量Jo2抗体腹腔注射SCID鼠,有利于人鼠嵌合肝中人肝细胞的增殖和存活。

BALB/c小鼠原位移植肝癌模型的建立及鉴定

目的：利用小鼠肝癌细胞株H22细胞肝脏原位种植法建立BALB／c小鼠原位移植肝癌模型，为研究肝脏肿瘤奠定基础。方法取新鲜小鼠腹腔传代的H22细胞，生理盐水洗涤2次，调细胞浓度为1×10^7／ml，与Matrigel按4∶1比例混合，用10μl微量进样针在开腹直视下沿肝叶长轴种植于BALB／c小鼠的肝左叶，每只5μl，逐层关腹，常规饲养2周后，处死小鼠观察肿瘤形成情况，检测小鼠肝脏原位癌的形成率及小鼠存活率，形态学和病理学方法鉴定造模效果。结果肉眼可见小鼠肝脏出现单个灰白肿瘤结节，病理学方法证实为肝癌，小鼠存活率和成瘤率均为95％，肿瘤大小均匀。结论成功建立了一种稳定、简单可行的BALB／c小鼠原位移植肝癌模型。