Cryopreservation is a crucial aspect of domestic animal ART facilitating valuable genetic propagation and preservation. However, despite improvements in cryopreservation, cryodamage is still an inevitable consequence due to the high lipid content in preimplantation embryos of these species. In additional, the mechanism of formation of lipid droplets (LDs) were not well understood. Herein, we propose a novel two-step delipidation (DeL) method then evaluate embryo viability and post- vitrification survival. Cleavage rate (84.3±8.3%) and blastocyst rate (30.6±8.8%) of delipidated zygotes were not different from the controls (80.3±4.1% and 29.8±9.9%) suggesting a valid technique for delipidated embryo production. Next, we identified smaller LDs forming into larger LDs at early embryo cell division (2-8 cell) and hypothesize a potential LDs fusion protein, CIDEA, is involved in LDs enlargement in bovine embryo. Our data provide a technique minimizing lipid content, and a further understanding of molecular pathway in LDs dynamic in bovine preimplantation embryo development.