Objective To use transperitoneal laparoendoscopic single-site (LESS) partial nephrectomy combined with flexible 3-D laparoscope for partial nephrectomy in pigs, and to introduce our experience on LESS training in a pig model for partial nephrectomy. Methods A male pig was given general anesthesia and was put in a supine position. A 3.5 cm incision was made through the right abdominal rectus. A multi-channel QuadPort(Olympus^TM) was established. Artificial pneumoperitoneum was created by 14 mmHg(1 mmHg=0.133 kPa); the lower pole and the kidney pedicle of the right kidney were isolated. The lower pole of the right kidney was resected after the renal artery was blocked. Figure-8 sutures were used to close the wound, with no notable bleeding noticed after opening the blood. The specimen was enclosed in an endoscopic pouch and pulled out from the incision. Results The procedure was smoothly completed without any extra incision. The operating time was 47 min; the period of renal artery occlusion was 21 min (5 min for resection, 16 min for stitching); and the intraoperative blood loss was 20 mL. There was no complication during the operation. Conclusion Our initial experience shows that 3D-LESS partial nephrectomy procedure is technically difficult, but it is safe, feasible and effective. Flexible 3D laparoscopy can provide clear 3D visualization, improving the operating accuracy and reducing the fighting of instruments. The pig model used in this study can help surgeons to obtain experience on 3D-LESS partial nephrectomy.