Objective?To study the anatomic characteristics of arcuate ligament and its clinical relevance as an anatomic landmark of the renal artery in retroperitoneal laparoscopic nephrectomy. Methods?297 patients were underwent retroperitoneal laparoscopic operations in our institute from Jan 2008 to July 2014. These included 186 cases of renal cancer, 74 cases of renal pelvis cancer and 37 cases of ureteral carcinoma. There were 176 male and 121 female patients with the average age of 54.6 years (range from 26 to 82 years). At first made a small incision in the lateral vertebral fascia, then dissociated the posterior pararenal space following the principle of “Dissociation according to need,Less rather than more”. The renal artery was found located at the psoas and the medial arcuate ligament of the diaphragm for anatomic landmark before the gap of the psoas.Then renal arteries were controlled by Hem-o-lok clips, the renal veins below the follow-up renal arteries were controlled in the same way. Results?The psoas major quadratus lumborum and diaphragm can be observed. The intersections of the diaphragm with psoas major and quadratus lumborum were bordered by the medial and lateral arcuate ligament, which looked like a seagull. The medial arcuate ligament extended towards the renal artery. This landmark enables us to locate renal artery quickly in all 297 cases. The operation time and risk of the vessel injury were reduced. Conclusions?The dissociation of the posterior pararenal space following the principle of “Dissociation according to need,Less rather than more” can reduce numbers of Trocar; The medial arcuate ligament is a significant anatomic landmark for dissecting renal artery during retroperitoneal laparoscopic nephrectomy. Under the positioning of the medial arcuate ligament is an effective way to locate renal artery, can reduce the operation time of retroperitoneal laparoscopic nephrectomy.