Effects of EGF and melatonin on gene expression of cumulus cells and furtherin vitroembryo development in bovines

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Summary: Despite previous research demonstrating the benefits of including growth factors and antioxidants to maturation medium to support embryo production, to date the effect of epidermal growth factor (EGF) and melatonin (Mel) on oocyte competency has not been studied. This study supplementedin vitromaturation (IVM) medium with EGF (10 ng/ml) and Mel (50 ng/ml) alone, or in combination, and evaluated cumulus cell (CC) gene expression and the development and quality of parthenogenetic blastocysts. No differences in CC gene expression levels indicative of developmental potential were found among the treatment groups. Antioxidant geneCuZnSODwas significantly (P< 0.05) decreased in CCs from the Mel group. Moreover, blastocyst rates on day 7 were significantly increased in EGF or Mel (P< 0.05), but not EGF+Mel. Significant decrease (P< 0.05) inGPX1,CuZnSOD,SLC2A1andHSPA1A(P= 0.07) mRNA levels was observed in blastocysts from the Mel group.OCT4gene expression was significantly increased (P< 0.05) in EGF+Mel and confirmed using immunofluorescence. Our results indicate that, despite the lack of changes of competence-related genes in CCs, IVM medium supplemented with Mel improved the culture environment sufficiently, resulting in improved blastocysts. Moreover, EGF and Mel combined during maturation increasedOCT4gene and protein expression in blastocysts, indicating its potential for stem cells.

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