Graphical summary. Here, the antioxidant capabilities of Mito-TEMPO induced blastocyst developmental competence through F-actin structure stabilization in bovine IVC embryos. Moreover, the Mito-TEMPO provided protection from cryo-injury in vitrified-warmed blastocysts, increasing the survival rate of blastocysts through direct mitochondria superoxide scavenger and F-actin stabilization in bovine embryos. Therefore, these findings contribute to the improvement of vitrification and warming procedures to overcome infertility.