Martinez Cristina A, Cuello Cristina, Parrilla Inmaculada, Maside Carolina, Ramis Guillermo, Cambra Josep M, Vazquez Juan M, Rodriguez-Martinez Heriberto, Gil Maria A, Martinez Emilio A
Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, International Excellence Campus for Higher Education and Research (CMN), University of Murcia, 30100 Murcia, Spain.
Department of Biomedical & Clinical Sciences (BKV), BKH/Obstetrics & Gynaecology, Faculty of Medicine and Health Sciences, Linköping University, SE-58185 Linköping, Sweden.
Antioxidants (Basel). 2022 Jun 15;11(6):1177. doi: 10.3390/antiox11061177.
Cloned and transgenic pigs are relevant human disease models and serve as potential donors for regenerative medicine and xenotransplantation. These technologies demand oocytes and embryos of good quality. However, the current protocols for in vitro production (IVP) of pig embryos give reduced blastocyst efficiency and embryo quality compared to in vivo controls. This is likely due to culture conditions jeopardizing embryonic homeostasis including the effect of reactive oxygen species (ROS) influence. In this study, the antioxidant melatonin (1 nM) in the maturation medium, fertilization medium, or both media was ineffective in enhancing fertilization or embryonic development parameters of in vitro fertilized oocytes. Supplementation of melatonin in the fertilization medium also had no effect on sperm function. In contrast, the addition of melatonin to the embryo culture medium accelerated the timing of embryonic development and increased the percentages of cleaved embryos and presumed zygotes that developed to the blastocyst stage. Furthermore, it increased the number of inner mass cells and the inner mass cell/total cell number ratio per blastocyst while increasing intracellular glutathione and reducing ROS and DNA damage levels in embryos. Contrarily, the addition of melatonin to the embryo culture medium had no evident effect on in vivo-derived embryos, including the developmental capacity and the quality of in vivo-derived 4-cell embryos or the percentage of genome-edited in vivo-derived zygotes achieving the blastocyst stage. In conclusion, exogenous melatonin in the embryo culture medium enhances the development and quality of in vitro-derived embryos but not in in vivo-derived embryos. Exogenous melatonin is thus recommended during embryo culture of oocytes matured and fertilized in vitro for improving porcine IVP efficiency.
克隆猪和转基因猪是相关的人类疾病模型,也是再生医学和异种移植的潜在供体。这些技术需要高质量的卵母细胞和胚胎。然而,与体内对照组相比,目前猪胚胎体外生产(IVP)方案的囊胚效率和胚胎质量较低。这可能是由于培养条件破坏了胚胎内环境稳定,包括活性氧(ROS)影响的作用。在本研究中,在成熟培养基、受精培养基或两种培养基中添加抗氧化剂褪黑素(1 nM),对提高体外受精卵母细胞的受精率或胚胎发育参数无效。在受精培养基中添加褪黑素对精子功能也没有影响。相反,在胚胎培养基中添加褪黑素可加速胚胎发育进程,增加发育至囊胚阶段的分裂胚胎和假定受精卵的百分比。此外,它增加了每个囊胚的内细胞团细胞数量和内细胞团细胞数与总细胞数的比例,同时增加了胚胎细胞内谷胱甘肽水平,降低了ROS水平和DNA损伤程度。相反,在胚胎培养基中添加褪黑素对体内来源的胚胎没有明显影响,包括体内来源的4细胞胚胎的发育能力和质量,或基因组编辑的体内来源受精卵发育至囊胚阶段的百分比。总之,胚胎培养基中的外源性褪黑素可提高体外来源胚胎的发育和质量,但对体内来源胚胎无效。因此,建议在体外成熟和受精的卵母细胞胚胎培养过程中添加外源性褪黑素,以提高猪IVP效率。