<p>Assissted oocyte activation (AOA) has emerged as a promising method to overcome fertilization failures that can occur after intracytoplasmic sperm injection (ICSI) due to the sperm's inability to adequately stimulate the oocyte. Fertilization failure after ICSI accounts for approximately 30% of human oocyte failures, contributing to an estimated total fertilization failure rate of 2–3%. However, concerns remain regarding the potential epigenetic changes that could influence both placental and fetal development. This study aims to examine the methylation and expression changes of key imprinted genes (DLK1 and MEG3) in umbilical cord blood and placental tissue. Therefore, the methylation and expression changes of DLK1 and MEG3 were compared among ICSI, ICSI-AOA, and natural fertilization groups. The analysis involved DNA methylation and real-time PCR. Results indicated no significant differences in overall methylation levels between the groups, although individual CpG positions displayed significant variations. Similarly, gene expression levels did not differ significantly across the groups. The study concludes that ICSI-AOA does not significantly impact the DNA methylation or gene expression of the imprinted genes (DLK1 and MEG3), suggesting that both ICSI and ICSI-AOA appear to be safe and reliable for infertility treatments. However, further research is essential to explore the long-term effects and safety profiles associated with ICSI-AOA.</p>