Background: With advances in diagnostic and therapeutic modalities, earlier detection of ectopic pregnancy with minimal symptoms makes laparoscopic management more common. However, if diagnostic delay occurs, significant hemoperitoneum associated with ectopic pregnancy is still a potentially life-threatening condition, which presents a therapeutic challenge especially when minimally invasive surgical approach is a significant concern. Herein, retrospective observational study in 18 consecutive cases of ectopic pregnancy with significant hemoperitoneum ≥ 800 mL was performed to assess the feasibility and efficacy of gasless laparoendoscopic single-site (LESS) surgery with intraoperative autologous blood cell salvage and donation for management. Results: At triage, median shock index was 0.95, median hemoglobin value was 6.5 g/dL, and median serum β-hCG value was 13,651 U/L. Surgical diagnosis included 7 ampullary (including 5 abortions and 2 ruptures), 5 ruptured isthmic (including 1 heterotopic pregnancy in remnant tube after salpingectomy and 1 remnant tube pregnancy after salpingo-oophorectomy), 4 ruptured interstitial (including 1 case after salpingectomy), and 2 ruptured ovarian pregnancies. Among these cases, 16 cases were successfully managed by LESS surgery, while, in 2 cases with interstitial pregnancy, conversion to multi-port laparoscopic surgery along with transfusion of bank blood were required. Laparotomic conversion was not experienced. Surgical procedures in 16 cases successfully completed by LESS surgery with intraoperative autologous blood transfusion included unilateral salpingectomy for 7 ampullary and 5 isthmic pregnancies, cornuotomy and cornual resection for each 1 case with interstitial pregnancy, and unilateral salpingooophorectomy and partial ovarian resection for each 1 case with ovarian pregnancy. Median surgical duration was 57.5 min, median amounts of salvaged peritoneal blood were 950 mL (range 800-2000 mL), and median amounts of transfused autologous blood were 540 mL (range 450-1300 mL). Major complications associated with LESS surgery and intraoperative autologous blood transfusion were not experienced. Conclusion: Gasless LESS surgery with intraoperative autologous blood transfusion is a feasible minimally invasive surgical option with avoidance to use homologous blood for management of selected cases of ectopic pregnancy even with significant hemoperitoneum.