Objective To use Spider system, a novel laparoendoscopic single-site (LESS) platform, for partial nephrectomy in pigs, so as to evaluate the feasibility and ergonomic features of the Spider system. Methods A 2 cm incision was made on the lateral border of the rectus muscle; the subcutaneous tissues and muscle layer were dissected bluntly by the Kelly clamp. With the outside sheath the Spider system was introduced into the peritoneal cavity. After establishment of pneumoperitoneum, a 5 mm scopy was introduced. The whole Spider platform was fixed using the suspension system on the bedside. The procedures of partial nephrectomy were performed according to the conventional steps. Renal reconstruction was done by the 2-0 V-loc barbed suture using the knotless closure technique. Results The procedure of partial nephrectomy was successfully performed in 2 cases, with the operation time being 100 and 102 min, the docking time being 7 and 11 min, and the time periods of warm ischemia being 23 and 28 min. The V-loc barbed suture was used to close the defect of kidney without bleeding after unclamping the renal artery. Conclusion This novel Spider LESS platform is safe and feasible for partial nephrectomy in pigs, which better solves the problem of “cross manipulation” in single-port laparoscopy, avoiding external instrument clashing; but it still has many drawbacks like the lack of strength and precise maneuverability of the tips, which may hamper the visual field in the operation.