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Selenium is commonly used as an antioxidant in a serum‐free culture medium setting. However, lycopene has emerged as a potent antioxidant being twice as efficient as β‐carotene and 10 times as efficient as α‐tocopherol with beneficial effects when supplemented in a serum‐free maturation medium. Here, we aimed to evaluate the effect of lycopene supplementation in a serum‐free culture medium on blastocyst development and quality. After in vitro maturation and fertilization, presumed zygotes were cultured in groups of 25 in 50 μl droplets of synthetic oviductal fluid. Culture medium supplementation was done using four experimental groups: insulin, transferrin, selenium (ITS, control); ITS + DMSO (diluent control); ITS + DMSO‐lycopene 0.1 μM (ITSL); and IT + DMSO‐lycopene 0.1 μM (ITL). DMSO was used as a diluent for lycopene. Blastocyst development among experimental groups was fitted in mixed‐effects models, and blastocyst quality parameters (assessed via differential apoptotic staining) were evaluated in mixed linear regression models. The cleavage (85.3 ± 2.4, 82.6 ± 2.7, 86 ± 2.3 and 86.4 ± 2.3% for control, diluent control, ITSL and ITL, respectively) and day 8 blastocyst rates (37.4 ± 3.3, 36.9 ± 3.4, 39.7 ± 3.3 and 46.2 ± 3.4% for control, diluent control, ITSL and ITL, respectively) were not different (p >.1) among experimental groups. Embryos produced in the ITL group resulted in blastocysts with higher total cell numbers (TCN; 141 ± 19.2), inner cell mass (ICM; 65.3 ± 11.6) and trophectoderm cells (TE; 75.2 ± 8.8) compared with the control (129 ± 19.2, 56.3 ± 11.6 and 72.7 ± 8.8, for TCN, ICM and TE; p <.01, respectively). Lycopene‐supplemented groups (ITSL and ITL) resulted in blastocysts with similar TCN, ICM and TE (p >.2). The number of apoptotic cells was not different among experimental groups (p >.1). Lycopene supplementation to the culture medium only produced a numerical increase in the blastocyst rate but replacing selenium with lycopene in a serum‐free culture medium resulted in blastocysts with more cells. [ABSTRACT FROM AUTHOR]