Effects of culture media and inhibitors on biology of porcine early embryonic development in vitro

Abstract

In vitro-production of porcine embryos is crucial for biomedical and agricultural research, however, current culture systems for porcine embryos are sub-optimal, and the developmental potential of in vitro-produced embryos is not well studied. The objectives of this study were to evaluate whether PZM-3 or NCSU-23 better support porcine embryo development and the effects of α-amanitin and cycloheximide on porcine embryonic development. Briefly, porcine presumptive zygotes were produced after in vitro maturation/fertilization (IVM/IVF) and cultured either in NCSU-23 or PZM-3 culture media. Transcript levels of BAX and BCL2L1 genes from blastocysts were detected by using Real-time PCR. The effects of α-amanitin and cycloheximide were evaluated for the role of inhibiting transcription and translation during early porcine embryogenesis. Results showed that both cleavage and blastocyst rates decreased significantly in NCSU-23 group compared as PZM-3 group. However, BAX and BCL2L1 transcript levels were similar in blastocysts cultured in both PZM-3 and NCSU-23 media. When porcine embryos cultured in PZM-3, cleavage rates were significantly decreased in the present of cycloheximide and both α-amanitin and cycloheximide treatments completely inhibited the blastocyst formation. Results showed that PZM-3 medium better supported porcine early embryonic development than NCSU-23 medium, and the inhibition of embryonic genome activation does not completely stop embryo cleavage but prevents development to the blastocyst stage.