Objective To establish a stable mouse model of limb heterotopic composite tissue allograft transplantation. Methods 25 pairs of C57BL/6 mice were used for this limb heterotopic allograft transplantation experiment. The hind legs were taken from the donor mice. A transverse incision was made at the level of inguinal ligament to expose and mobilize the femoral vessels up to the external iliac vessels. All branches to the external iliac artery and vein were cut off by electrocoagulation. To cut off the external iliac artery and external iliac vein, and to transect the femur, thigh muscles and skin. The donor external iliac artery-external iliac vein anastomosis was performed in end-to-side fashion with recipient mice's right common carotid artery and jugular vein, respectively. Continuously suture the donor skin to the recipient skin incisal margin to fix the limb. Cut off the donor limb's distal tibial joint, and ligate the stump. Results The success rate of the heterotopic limb transplantation operation was 76%. The total operation time was 87.1±10.1 minutes, and the vessel anastomosis time was 55.3±9.6 minutes. Conclusions This mouse model of limb heterotopic limbcomposite tissue allograft transplantation shows a high success rate of the surgical operation, and is stable and reliable. It is a new ideal model for studies on transplantation immunology.