원문정보
초록
영어
In vivo oocytes grow and mature in ovarian follicles whereas oocytes are matured in vitro in plastic culture dishes with a hard surface. In vivo oocytes show a superior developmental ability to in vitro counterparts, indicating suboptimal environments of in vitro culture. This study aimed to evaluate the influence of an agarose matrix as a culture substrate during in vitro maturation (IVM) on the development of pig oocytes derived from small antral follicles (SAFs). Cumulusoocyte complexes (COCs) retrieved from SAFs were grown in a plastic culture dish without an agarose matrix and then cultured for maturation in a plastic dish coated without (control) or with a 1% or 2% (w/v) agarose hydrogel. Then, the effect of the soft agarose matrix on oocyte maturation and embryonic development was assessed by analyzing intra-oocyte contents of glutathione (GSH) and reactive oxygen species (ROS), expression of VEGFA, HIF1A , and PFKP genes, and blastocyst formation after parthenogenesis. IVM of pig COCs on a 1% (w/v) agarose matrix showed a significantly higher blastocyst formation, intra-oocyte GSH contents, and transcript abundance of VEGFA. Moreover, a significantly lower intra-oocyte ROS content was detected in oocytes matured on the 1% and 2% (w/v) agarose matrices than in control. Our results demonstrated that IVM of SAFs-derived pig oocytes on a soft agarose matrix enhanced developmental ability by improving the cytoplasmic maturation of oocytes through redox balancing and regulation of gene expression.
목차
INTRODUCTION
MATERIALS AND METHODS
Oocyte collection and in vitro growth (IVG)
Preparation of an agarose matrix
IVM
PA and in vitro culture (IVC) of PA embryos
Measurement of intra-oocyte GSH and ROS contents
Quantitative real-time polymerase chain reaction (RT-qPCR)
Statistical analysis
RESULTS
Effects of agarose matrix on oocyte maturation and blastocyst formation
Effect of agarose matrix on intra-oocyte GSH and ROS contents of IVM oocytes
Effects of agarose matrix on gene expression of CCs
DISCUSSION
CONCLUSION
REFERENCES