earticle

영어

The success of in vitro embryo production (IVP) demonstrates that it’s possible to bypass the oviduct throughout early development. However, several studies show that embryos developed in vivo are superior to embryos developed in vitro. Using an ex vivo model of porcine uterus is one of the strategies to investigate fertilization within the oviductal environment. During this study, in vitro matured porcine oocytes (MII) were fertilized with 7.5×107, 15×107 and 30×107 sperm cell for 20 min in porcine uterine ex vivo model. The oocytes were then flushed and performed in vitro culture (IVC) at 39°C for 168 hours under 5% O2, 5% CO2. MII oocytes used for in vitro fertilization (IVF) served as control-1. Before IVF, MII oocytes cultured in porcine uterine ex vivo model for 20 min served as control-2. Within the results, penetration rate, MPN formation, monospermy, polyspermy, and efficiency of fertilization had not shown significant difference between control-1 and control-2 group, respectively. However, penetration rate (treatments: 29.7±4.4, 34.3±3.2, 44.3±7.4 vs. 80.0±1.7), polyspermy (treatments: 5.7±5.7, 9.7±5.8, 8.0±4.0 vs. 33.7±9.5) and efficiency of fertilization (treatments: 23.7±2.3, 29.0±3.6, 35.0±4.6 vs. 43.0±5.8) were significantly decreased in treatment groups compared to control-1 (p<0.05). GSH accumulated levels were significantly decreased in 30×107 sperm cell treated group compared to control-1 (p<0.05) and there was no significant difference in ROS accumulated levels among the groups. For embryo development, the cleavage rate and blastocyst rate had not shown significant difference between control-1 and control-2 group. However, the cleavage rate (treatments: 16.3±2.6, 20.1±2.7, 40.7±13.4 vs. 69.5±6.3, 74.2±3.4) was significantly decreased in treatment groups compared to control-1 and control-2 (p<0.05). And the cleavage rate in the treatment group of 30×107 (40.7±13.4) was significantly higher than the treatment group of 7.5×107 (16.3±2.6) (p<0.05). The blastocyst rate (treatments: 31.7±4.0, 25.7±4.0, 26.7±6.5 vs. 7.2±2.4, 9.9±3.0) was significantly increased in control-1, control-2 and the treatment group of 30×107 compared to 7.5×107 and 15×107 (p<0.05). Therefore, these results suggest that ex vivo model may decrease the penetration rate and efficiency of fertilization by reducing GSH accumulated levels. Cleavage rate and blastocyst rate can be promoted by increasing sperm number during ex vivo fertilization.