earticle

영어

2i system is useful for maintenance of navie state of embryonic stem cells in various species. However, the role of 2i system in porcine preimplantation stage embryos unclear. In this study, we investigated the quality of blastocyst and pluripotent-related factor by 2i system. And, we also investigated about DNA and RNA epigenetic modification status by 2i system during porcine early embryo development. After treatment of MEK1/2 inhibitor PD0325901(4 μM), GSK-3 inhibitor CHIR99021(0.3 μM) and 2i (PD0325901+CHIR99021) during pre-implantation development, blastocyst formation rate had no significantly difference. However, blastocyst size increased in 2i treatment embryos. Blastocyst quality also increased after treatment of CHIR and 2i using TUNEL assay and Brdu assay. Interestingly, main transcription factor OCT4 and SOX2 positive cells in blastocyst was significantly increased in 2i groups. Especially, histone modification- relation proteins like h3k9me3 and h3k9ac changed after treatment of CHIR and 2i. Global gene expression patterns related with pluripotency and epigenetic also changed in inhibitor treatment groups. Furthermore, bi-sulfite sequencing results showed that 2i treatment groups demethylated in satellite I region. In conclusion, our findings strongly suggest that 2i system can be useful method for increasing of blastocyst quality by affecting of ICM/TE formation by regulating epigenetic modification during early embryo development.