ObjectiveTo explore the surgical techniques of rat orthotopic liver transplantation (OLT) model in rats using modified technique by single operator. MethodsA total of two hundreds cases of rats (one hundred pairs) were used to establish OLT model including eighty cases of rats (forty pairs) used in formal study by using modified Kamada “two-cuff” method, of which the key surgical procedures were further modified. Intraoperative results and postoperative recovery were analyzed. ResultsAll eighty cases of rats successfully operated. Operative time in donors was (28.5±2.4) min and reparative time of donor liver was (10.2±1.8) min. Anastomosis time for suprahepatic inferior vena cava and portal vein (15.3±1.9) min and (3.4±1.2) min, respectively. Anhepatic phase of transplant recipients was (23.8±1.9) min, anastomosis time for infrahepatic inferior vena cava was (5.1±2.1) min, and biliary reconstruction time was (3.1±0.9) min. Blood loss of donor and recipients during the operation was blew 0.5 ml. No recipient died for operation. The 2 d survival rate of recipients was 90% (36/40), three rats died of bleeding at the seam in hepatic inferior vena cava and one rat died of longer anhepatic phase. The 7 d survival rate of recipients was 82.5% (33/40), three rats died of intra-abdominal infections. ConclusionThe modification in en bloc separation of abdominal aorta and interior vena cava for abdominal aortic perfusion of donor liver, careful dissection of left subphrenic vein triangle and the hemostasis by heat coagulating can reduce operative complications, guarantee the quality of donor liver and improve the survival rate of rats in establishment of OLT model in rats using modified technique by single operator.