Authors : Se-Heon Jeung, Yu-Byeol Jeon , Dibyendu Biswas , Kyung-Chul Choi , Eui-Bae Jeung and Sang-Hwan Hyun
Abstract: Porcine In Vitro Maturation (IVM) and In Vitro Fertilization (IVF) technique has been used and improved. However, polyspermic penetration, low rate of Blastocyst (BL) formation and poor quality of BLs are induced by imperfect nuclear and cytoplasmic maturation. It has been reported that Epidermal Growth Factor (EGF) is beneficial for oocyte maturation to improve the IVM system and Amphiregulin (AREG) is a growth factor containing an EGF-like domain. Consequently, the present study was performed to investigate whether during porcine IVM system EGF and/or AREG supplementation is profitable for improving oocyte maturation and embryo development. In Experiment 1, oocytes were maturated with EGF (15 ng mL-1) or AREG (15 ng mL-1) or combination of both and after 44 h, oocytes were stained with Hoechst and metaphase II and development rate were evaluated. Without any growth factor served as a control. In Experiment 2, maturated oocytes from the previous groups were fertilized with proven sperm and penetration rate after 10 h of post-insemination was evaluated. In Experiment 3 in vitro matured porcine oocytes were stained with fluorescein isothiocyanate-labeled peanut agglutinin and the distribution of Cortical Granules (CGs) was evaluated by laser confocal microscopy. In Experiment 1, the oocytes maturation rate were significantly higher in EGF, AREG or combined group (70.9±15.5, 75.8±9.8 and 70.5±10.8%, respectively) compared to control (51.6±18.3%) but no significant difference among the treatments group. In Experiment 2, there was no significance of sperm penetration rate among the groups however, polyspermy and sperm number per oocytes were decreased significantly (p<0.05) in combined group (26.7±12.0 and 1.35±0.2%, respectively) compared with control (43.5±13.1 and 1.63±0.3%, respectively). In Experiment 3, BL formation rate were significantly (p<0.05) higher in combined group (17.1±5.0%) compared with control and EGF group (9.4±3.2 and 13.1±4.6%, respectively) but the TE cell number and total cell number were significantly higher in (p<0.05) combined group compared to control and other treatment group. In Experiment 3, the CG area was increased significantly (p<0.05) in all treatment groups compared with control group (11.1±3.4, 9.7±3.0 and 10.4±2.8 vs. 4.1±1.8%, respectively). These results indicate that the addition of EGF and/or AREG to porcine IVM medium that enhances oocyte maturation and increases the total cell number. Combination of both growth factors also increases development rate and decreases polyspermy. Therefore, it is suggested that AREG can assist immature porcine oocytes to the metaphase II stage, enhance developmental potential in in vitro system and that there is due to synergistic effect of EGF and AREG during oocytes and embryonic development.
Se-Heon Jeung, Yu-Byeol Jeon , Dibyendu Biswas , Kyung-Chul Choi , Eui-Bae Jeung and Sang-Hwan Hyun , 2012. Effect of EGF and AREG Treatment During Porcine in vitro Maturation on in vitro Developmental Potential of Preimplantation Embryos. Journal of Animal and Veterinary Advances, 11: 1100-1105.