This study was designed to investigate whether acteoside (antioxidant) improves canine SCNT efficiency. Cell cycle of canine fetal fibroblasts was synchronized by culturing to contact inhibition, serum starvation and treating with acteoside. Cell cycle synchronization, apoptosis and reactive oxygen species (ROS) detection were analyzed using flow cytometry. The fibroblasts, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were reconstructed in enucleated oocytes. Embryos using acteoside-treated cell were surgically transferred into oviducts of estrus cycle synchronized recipient dogs. Based on flow cytometry, there was no significant difference between serum starvation (88.2%), contact inhibition (84.6%) and most acteoside groups (82.4384.48%, P>0.05). The rate of ROS and apoptosis in acteoside group (42.75 and 4.17%) showed significantly decrease between contact inhibition (54.33 and 10.63%) and serum starvation (99.5 and 62.37%). After SCNT, fusion rate of acteoside group was 66.7%, which was higher than those of contact inhibition (52.5%). Developmental rate to the 8 cell and 10 cell stages was higher in acteoside-treated groups (14.8 and 4%) than those of contact inhibition (5 and 0%). Total 38 SCNT embryos using acteoside-treated fibroblasts were transferred into three recipient dogs and one recipient finally delivered one puppy. In conclusion, this study demonstrated that canine fibroblasts could be arrested at the G0/G1 stage with reduced ROS and apoptosis after acteoside treatment. And reconstructed canine embryos with acteoside-treated cells forwarded subsequent embryo development. These results may contribute to improve the efficiency of canine SCNT. This research was supported by Cooperative Research Program for Agriculture Science & Technology Development (No. PJ009333), Rural Development Administration, Republic of Korea.
02 - 04 Sep 2014
World Congress of Reproductive Biology