An antifreeze protein from Anatolica polita (ApAFP914) was tested on sheep embryos to improve their survival after freezing and thawing. The study compared slow freezing and vitrification methods for freezing sheep oocytes fertilized in vitro. Adding different concentrations of ApAFP914 to the vitrification liquid showed higher survival and hatching rates for sheep embryos compared to slow freezing. Embryos at the expanded blastocyst stage had the best survival and hatching rates. Fast-growing embryos had higher rates than slow-growing ones. Adding 10 μg/mL of ApAFP914 increased the hatching rate of slow-growing embryos. Vitrification was found to be more effective than slow freezing for preserving sheep embryos in vitro, and using ApAFP914 could boost the hatching rate of embryos after cryopreservation.