Objective To perform partial nephrectomy via combined transgastric and transvesical approach in porcine and to use V-loc suture for suturing the renal defect, so as to further assess the safety and feasibility of combined transgastric and transvesical approach. Methods Three female pigs were used in this study. The transvesical access was established by incising the anterior wall of the bladder under ureteroscope, and a self-designed Trocar was inserted. Under the guidance of ureteroscopy, the transgastric access was established by a needle knife with cautery, and the gastroscope was introduced after balloon dilation. Under the guidance of gastroscope, laparoscopic ultrasonic scalpel was introduced via the bladder Trocar; the kidney artery and vein were separated, the arteries were blocked and some lower part of renal parenchyma was resected; and the wound was closed by 2-0 V-loc suture which was introduced percutaneously. The specimens were collected from the bladder. Results Six partial nephrectomy procedures were done in the 3 pigs. The first case was transferred to traditional laparoscopy due to incompleted blocking of the artery, bleeding and loss of vision. The rest 5 cases were successfully completed, with a mean operation time of 174 min(140-220 min), a warm ischemia time of 21 min (17-28 min), and with no significant bleeding. The average size of incised tissue was 1.5 cm×1.5 cm. Conclusion Partial nephrectomy via combined transgastric and transvesical approach is extremely difficult, but it can be done by using the V-loc suture and auxiliary needle laparoscopic instrument; however, its clinical prospect still needs further verification.