To assess the effects of estradiol (E2) on trigger day on cumulative live birth rates (CLBRs), and pregnancy outcomes after fresh and frozen-thawed embryo transfer (FET).This multicenter retrospective cohort study included 42 315 patients from five reproductive centers. Six subgroups were divided according to E2 on trigger day (<1000, 1000-2000, 2000-3000, 3000-4000, 4000-5000, >5000 pg/mL). Smooth curve fitting and nonlinear mixed-effects models were used.When E2 was <5500 pg/mL, the CLBR increased by 10% for every 1000 pg/mL increase in E2. When E2 was between 5500 and 13 281 pg/mL, CLBR increased by 1.8% for every 1000 pg/mL increase in E2. When E2 was >13 281 pg/mL, CLBR decreased by 3% for every 1000 pg/mL increase in E2. From group E2 < 1000 to group E2 > 5000 pg/mL, pregnancy and live birth rates in fresh cycles were not related to E2. The live birth rate after FET was higher in the E2 ≥ 5000 pg/mL group than in the E2 < 1000 pg/mL group (odds ratio [OR] 4.03, and 95% confidence interval [CI] 3.74-4.35; adjusted OR 1.20, 95% CI 1.05-1.37).CLBR is associated with E2 on trigger day in a segmented manner. Pregnancy and live birth rates in fresh cycles were not associated with E2. The live birth rate in FET cycles was highest when E2 ≥ 5000 pg/mL.© 2023 International Federation of Gynecology and Obstetrics.