Effects of amino acids on maturation, fertilization and embryo development of pig follicular oocytes in two IVM media

Abstract

This study was conducted to develop a serum-free, defined medium for IVM of pig oocytes. Modified North Carolina State University (mNCSU)-23 media with or without supplementation with both epidermal growth factor (EGF) and gonadotrophin were used as base media. In separate experiments, each base medium was supplemented with porcine follicular fluid (pFF), polyvinyl alcohol (PVA), PVA and essential amino acids (EAA), PVA and nonessential amino acids (NEAA) or PVA with both EAA and NEAA. Averaged across these five treatments, the percentage of blastocyst formation was higher ( P<0.05) in the base medium supplemented with EGF and gonadotrophins. In both base media, the addition of NEAA yielded similar percentages of maturation (81–82% versus 75–80%), sperm penetration (89–93% versus 80–86%) and blastocyst formation (4–18% versus 4–13%) as media supplemented with pFF. Although similar benefits were found after the addition of EAA, their addition was associated with lower ( P<0.05) maturation (66%) and sperm penetration (58%) than when pFF was added to the base medium without EGF and gonadotrophins. However, decreased maturation after EAA addition was not detected in the base medium containing EGF and gonadotrophins. Within the same base medium, monospermy, male pronucleus formation, cleavage and blastocyst formation were not affected by the treatments; and combined addition of EAA and NEAA did not further improve oocyte development. In conclusion, a maturation system using a defined mNCSU-23 medium supplemented with EGF, gonadotrophins and EAA or NEAA was developed which yielded a similar number of blastocysts compared with a pFF-containing medium.