Different Methods Of Parthenogenetic Activation On Porcine Oocytes Research

In this study, porcine oocytes in vitro maturation, as experimental material, were used by single electrical parthenogenetic activation, the twice electrical parthenogenetic activation, chemical parthenogenetic activation and physical methods combined with chemical reagents parthenogenetic activation, and analyzed the effects of various methods of activation. The experiment was conducted to choose a good parthenogenetic activation method to improve the cleavage rate and blastocyst rate after parthenogenetic activation of porcine oocytes in vitro maturation, so as to improve the success rate of nuclear transfer, as well as to lay the foundation for further study.This study includes the following main elements: 1. The different electric field intensity and pulse duration was choosed in single parthenogenetic electrical activation to compare the efficiency of different activation combinations. 2. The pulse duration and pulse frequency was fixed, different electric field strength was choosed respectively in twice electrical parthenogenetic activation to compare early parthenogenetic embryo development impact of the different combinations. 3. Ionomycin with different concentrations was used for different time; Ionomycin was combined with 6-DMAP, CB and CHX respectively; 9% Ethanol was used for different time. The efficiency of each combination was compared to study the better chemical parthenogenetic activation method. 4. Electrical activation treatment was combined with chemical reagents(6-DMAP, CB and CHX), then different combinations of pairs were compared to research the early developmental effects after parthenogenetic activation of oocytes in vitro maturation.The results showed that:1. To determine the single electrical parthenogenetic activation parameters of porcine oocytes in vitro maturation, the best condition was 2.0kV/cm, 1DC, 10μs, 20μs or 30μs, and 30μs was ideal;2. For twice electrical parthenogenetic activation parameters of the porcine in vitro maturation oocytes, the best condition was 100 kV/cm, 1.4kV/cm, 30μs, 1DC;3. In the study of the effect of Ionomycin for different time with different concentrations in porcine oocytes parthenogenetic activation, the better conditions were 20mmol/L Ionomycin for 10min or 15min, and the most ideal conditions is for 10min;4. The cleavage rate and blastocyst rate of groups after parthenogenetic activation with Ionomycin combined with 6-DMAP, CB respectively was higher than the group with Ionomycin and CHX;5. The cleavage rate and blastocyst rate after parthenogenetic activation of 9% Ethanol for 10min, 20min were significantly higher than that for 30min, 40min, and there was no significant difference between the former two groups;6. The efficiency of the conditions with 6-DMAP or CB after electrical parthenogenetic activation was significantly better than that of the conditions with CHX.