The study for development of in vitro culture system of mammalian oocyte by global analysis of cumulus cell-stimulating substrate during ovulating process

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During ovulation phase, granulosa cells-derived secretion factors stimulate cumulus cells, which result in completion of oocyte maturation. Since current culture system of oocyte employs only cultivation of cumulus-oocyte complexes(COCs), rate of oocyte maturation is low compared with that of oocyte collected from in vivo due to lack or insufficient of granulosa cells-derived oocyte maturation factors. In this research, we showed that TACE/ADAM17 expressed in granulosa cells and cumulus cells cleaved EGF-like factors from granulosa cells and cumulus cells, which resulted in induction of oocyte maturation via EGFR-ERK1/2 pathway in cumulus cells during ovulation process. We also investigated that PGE2-PTGER2, 4 pathways enhanced expressions of TACE/ADAM17 and EGF-like factors, thereby sustained EGFR-ERK1/2 pathway in cumulus cells during oocyte maturation. Furthermore, c-Src phosphorylated by FSH-PKC stimuli induced association with TACE/ADAM17, which required for protease activity of TACE/ADAM17 in granulosa cells and cumulus cells. Finally, optimization of expression and activation pattern of TACE/ADAM17 in cumulus cells of COCs by PGE2 or FSH-c-Src improved maturation rate of oocyte during in vitro maturation of porcine COCs