Glycine-L-Proline Media Supplementation Reduces Reactive Oxygen Species and Improves Preimplantation Development in Mouse Embryos
DOI:
https://doi.org/10.11113/mjfas.v22n2.4710Keywords:
Assisted Reproductive Technology, Embryo Development, Glycine, L-Proline, Mitochondria, Reactive Oxygen SpeciesAbstract
In vitro embryo culture systems often expose embryos to elevated oxidative stress levels, compromising development. Glycine (Gly) and L-Proline (L-Pro), naturally occurring amino acids with antioxidant properties, have been shown to mitigate reactive oxygen species (ROS). This study aimed to assess the individual and combined effects of Gly and L-Pro supplementation on preimplantation development in mouse embryos. Two-cell stage BALB/C mouse embryos were cultured in M16 medium supplemented with either Gly or L-Pro at concentrations of 0.2, 0.4, and 0.6 mM, and a combined group of 0.6 mM Gly + 0.6 mM L-Pro. Developmental progress to the 8-cell, morula, and blastocyst stages was recorded. Mitochondrial activity and ROS levels were quantified using MitoTracker Red and H2DCFDA assays, respectively. The combined treatment group exhibited significantly higher rates of normal morphology (94.3% 8-cell, 85.7% morula, 72.9% blastocyst) and developmental competence (92.9% blastocyst formation, 55.7% hatching) compared to controls (p < 0.01). ROS levels were lowest in the combined group (22.24 ± 1.54 pixel x10^5) and mitochondrial intensities were highest (105.1 ± 5.83 pixel x10^5), indicating improved mitochondrial activity and reduced oxidative stress. These findings suggest a complementary interaction between Gly and L-Pro in enhancing embryo quality and development by modulating oxidative stress and mitochondrial function. This novel supplementation strategy could inform future improvements in assisted reproductive technologies.
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