Background: It remains unclear whether patients undergoing three-dimensional (3D) laparoscopy-assisted gastrectomy could benefit from enhanced recovery protocol. The aim of present study is to compare enhanced recovery protocol and the conventional perioperative management after 3D laparoscopy-assisted gastrectomy in gastric cancer.Methods: A prospective cohort of patients received 3D laparoscopy-assisted gastrectomy were enrolled between 2017 and 2018. A hospital-based enhanced recovery protocol was established and implemented. Patient’s clinic-pathological characteristics and perioperative outcomes were compared between enhanced recovery group (ERG) and conventional group (CG). ER protocol compliance rate was calculated for patients in ERG. Univariate and multivariate binary logistic regression analysis were performed to investigate influential factors for delayed discharge and for postoperative complications.Results: One hundred and eighty-seven patients received 3D laparoscopy-assisted gastrectomy in ERG and 111 patients in CG were enrolled in the final analysis. Patients had comparable baseline characteristic between groups. However, patients in ERG had shorter time to oral feeding, reduced postoperative hospital stay and less medical cost (all P < 0.05). The postoperative complication rate were 10.7% for ERG and 10.8% for CG respectively. Regarding individual items in enhanced recovery protocols, prevention of postoperative nausea and vomiting had the highest compliance of 100% (187/187) while ambulation on postoperative day one had the lowest compliance of 32.1% (60/187). Univariate logistic regression analysis revealed operation time (P < 0.001), blood loss (P = 0.007), intraoperative transfusion (P = 0.003) and compliance (P < 0.001) were correlated with delayed discharge, while multivariate analysis demonstrated that only compliance [odds ratio (OR), 0.939; P < 0.001] and operation time (OR, 1.010; P = 0.048) were statistically significant. Additionally, univariate analysis showed blood loss (OR, 1.002; P = 0.028) and compliance (OR, 0.978; P = 0.030) were associated with postoperative complications, but multivariate analysis showed neither was statistically significant. Spearman correlation analysis revealed compliance was negatively correlated with postoperative hospital stay (Spearman r = -0.64, P < 0.001) and with medical cost (Spearman r = -0.26, P < 0.001).Conclusion: The present prospective cohort study suggests it is safe and feasible to incorporate 3D laparoscopic gastrectomy into enhanced recovery settings. Furthermore, improving compliance with enhanced recovery protocol may shorten hospital stay and promote postoperative recovery.