Evaporative-drying is one alternative to cryopreservation. It has the advantages that during sample preparation it does not require liquid nitrogen and a freeze-dryer machine; however, the results have not been as successful as freeze-drying. The objectives of this study were to determine whether artificial activation (electrical and chemical activation) could improve the development of porcine embryos in vitro after intracytoplasmic injected with evaporative-dried sperm, to optimize the activation protocol for porcine ICSI. The results show that, cleavage and blastocyst rate were significantly improved by oocytes activated with ionomycin for 5 min followed by treatment with 6-DMAP for 4 h (81.0 and 17.4%) or with an output voltage of 1.1 KV/cm for 30 μs (85.5 and 24.0%) after ICSI. When the oocytes activated with combination of chemical and electrical activation significantly increased activation, pronuclear formation, and cleavage, blastocyst formation rates. The diploidy of blastocysts was no significant difference in ICSI groups. Finally, we found that total apoptosis, fragmentation and TUNEL index of blastocysts were significantly decreased, the total cell number was significantly increased and ROS level did not increased when the oocytes underwent artificial activation stimuli. These results collectively indicate that artificial activation after ICSI is effective in elevating the embryos development, and improves the quality of blastocysts, while ensuring normal chromosome composition. Our findings suggest that artificial activation might contribute to the optimization of techniques for in vitro production of porcine embryos. This study was supported by a grant from the Next-Generation BioGreen 21 program (No.PJ009624) funded by Rural Development Administration, Republic of Korea.
02 - 04 Sep 2014
World Congress of Reproductive Biology