Objective：To investigate the surgical procedures for establishing a reliable murine model of heterotopic small bowel transplantation (SBTx), with the purpose of providing a good experimental tool for the rejective reaction study in SBTx. Methods：C57BL/6 mice were used as donor and the BALB/c mice used as recipient animals. The procedure of segmental heterotopic SBTx in allogenic mice was performed as following: the donor′s portal vein was anastomosed end-to-side to the recipient′s inferior vena cave (IVC), and then the donor′s superior mesenteric artery (SMA) with an aortic patch was anastomosed end-to-side to the recipient abdominal aorta. Subsequently, proximal end of the graft intestine was ligated and the distant end was anastomosed end-to-side to the recipient′s jejunum. The mice were fasted for 3 days, but free access to water after the operation. In addition, the mice were given 5% glucose-saline solution (2 mL) twice daily by subcutaneous injection, but without use of antibiotic and immunosuppressive therapy. The models survived beyond 5 days were considered to be successful. Results：Segmental SBTx were perfomed in 30 mice, with a postoperative day (POD) 5 survival rate of 70 % (21/30). The operating time in donor was (41±5.5) min, the graft warm ischemic time was about 0.5 min, the time for trimming the aortic patch in SMA was 3 min and cold preservation of donor′s small bowel was (30±7.5) min. The time of recipient preparation and transplantation was (90±7.5) min, in which the abdominal aorta and IVC blockage time was (40±3.0) min, the average time for the vein anastomosis and the arterial anastomosis was (10±2.0) min and (15±2.5) min, respectively. The average bleeding volume was 0.2 mL. The operation failed in 9 cases, due to arterial anastomotic thrombosis and anastomosis stenosis (6 cases), massive hemorrhage (2 cases) and intraperitoneal infection (1 case). Conclusions：The high quality of the donor′s small bowel segment harvesting, excellent vascular and intestinal anastomosis as well as adequate supplement of liquid to both donor and recipient are all crucial factors to ensure the efficiency and success of SBTx.