ABSTRACT
Background: The success of in vitro-fertilization (IVF) cycles is determined in large part by the quality of embryo cleavage, which in turn, is dependent on the quality of the embryo culture media (CM). Many factors can influence the quality of embryo CM, one of which is the levels of Cell Free Deoxyribonucleic acid (DNA). Understanding the association between Cell-free DNA levels in embryo CM and the quality of embryo cleavage could help improve the quality of IVF techniques. Methods: This prospective study was conducted with 96 spent CM from patients undergoing IVF cycle, in order to determine relationships of Cell-free DNA levels in embryo CM with embryo cleavage quality on day 3. After intracytoplasmic sperm injection (ICSI), 48 embryos were evaluated on day 3 of their development, according to their cell number. Day 2 and day 3 CM corresponding to each one of the embryos was analyzed, by quantitative PCR, for estimation of Cell-free DNA levels. Results: The results revealed a significant increase in Cell-free DNA levels on day 2 CM corresponding to 4 to 6 cell embryos compared to those corresponding to 7 to 8 cell embryos (p=0.04). As for day 3 CM, the results showed no significant difference between the Cell-Free DNA levels in CM of 7-8 and those of 4-6 cell embryos (p=0.4). Also, cell free DNA levels in embryo CM, were significantly higher on day 2 compared to day 3 for both 7-8 and 4-6 cell embryos (p=0.03; p=0.04). Conclusion: We conclude that cell-free DNA levels in CM might be associated with delayed embryo cleavage.
ABSTRACT
Background: The process of preimplantation embryo development in vitro represents a key phase during in vitro fertilization (IVF) cycles. It involves several regulatory signaling pathways as well as an optimized in vitro culture system. The resulting embryo quality helps to determine embryo competence before implantation and pregnancy outcomes. Reactive oxygen species (ROS) are known to play a major role in influencing the process of embryo development. Their role can be reflected in the regulation of signaling pathways as second messengers or in the irreversible cell alterations due to oxidative stress following an excess of ROS levels. Methods: In this study, we investigated the association between morphological embryo quality (fertilization, cleavage quality, and fragmentation levels) and lipid peroxidation levels (Malondialdehyde) in embryo culture media. After intracytoplasmic sperm injection (ICSI), a total of 103 oocytes were evaluated on day 1 and day 3 of their development, and their corresponding culture media were analyzed by estimating MDA levels using thiobarbituric acid. Results: The results showed no significant association between MDA levels in culture media and fertilization rate (p=0.3), sperm quality (p=0.99; p= 0.17; p=0.46; p=0.30; p=0.65; p=0.44; p=0.09; p=0.15; p=0.56), embryo fragmentation levels (p=0.79; p=0.40), AMH levels (p=0.31; p=0.36) and female age (p=0.60; p=0.34). However, we revealed a significant association between cleavage quality and MDA levels in the embryo environment (p=0.03). Conclusion: We conclude that oxidative stress in IVF culture media might be mainly associated with delayed embryonic development.