Background In our previous study, transumbilical endoscopic submucosal dissection (TU-ESD) was revealed to be feasible, but delayed gastric perforation was observed in 30% of ESD sites. In this study, we aimed to verify locations at which it is feasible to perform TU-ESD in the upper gastric body and to demonstrate the safety of TU-ESD in single-basin lymph node dissection (SBLND). Methods In vitro, TU-ESD was performed at three lesion sites (anterior wall, AW; posterior wall, PW; and lesser curvature, LC) in each porcine stomach using an EASIE-R tray (cases = 10). In vivo, TU-ESD was performed with SBLND in 9 pigs. Seven days after the operation, the pigs were sacrificed and examined. Results In the in vitro feasibility study, the TU-ESD time was significantly faster in the PW group (5.9 ± 2.0 min) than in the LC group (8.5 ± 1.5 min) (p < 0.05) in all 10 cases. In the in vivo survival study, TU-ESD with SBLND was successfully performed without any complications (N = 9). There were no cases of delayed perforation, and healing ulcers were found in all pigs 7 days after the operation. Ulcer size (5.2 ± 3.5 cm 2 ) was approximately 36% smaller than that observed at the ESD operation site (8.1 ± 1.9 cm 2 ) (p = 0.05). Epithelialization in the margin and healing of the gastric ulcers were confirmed by microscopy.Conclusions TU-ESD with SBLND is a feasible and safe method. The upper posterior gastric body could be the most feasible location for performing TU-ESD, perhaps because of the difference in the subcutaneous dissection time.Keywords Endoscopic submucosal dissection · Single-port surgery · Gastric neoplasm · Endoscope · Laparoscopy · Porcine Although the incidence and mortality rates of gastric cancer have gradually decreased in northeast Asia, gastric cancer remains the fifth most common malignancy and the third leading cause of cancer-related deaths worldwide [1,2].The prevalence of early gastric cancer (EGC) is higher than 50% in northeast Asia because of the recently implemented screening system and advances in endoscopic diagnosis [3]. The most common site of gastric cancer is the lower onethird of the stomach, in which over half of all such cases occur in Korea; however, the proportion of recent cases of gastric cancer occurring in the upper one-third of the and Other Interventional Techniques Sang-Ho Jeong and Jae-Seok Min contributed equally to this study.Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.