| Part I Study on the Effect of Activation on human unfertilized oocytes after intracytoplasmic sperm injectionObjectiveTo investigate the effect of different activation methods on human unfertilized oocytes after intracytoplasmic sperm injection (ICSI).DesignOocytes allocated to this study were those that did not show any signs of normal fertilization (extrusion of the second polar body, pronuclei formation) after ICSI for 24 hours, and that were still with no signs of degeneration or fragmentation. 200 oocytes were randomly subjected to one of five activation treatment, ethanol(ET), ethanol + 6-dimethylaminopurine (6-DMAP), ionomycin ( IO ) , ionomycin + 6-dimethylaminopurine (6-DMAP), 6-dimethylaminopurine (6-DMAP). Fluorescence in situ hybridization (FISH) was employed to analyze chromosomes using X and Y dual-colored probes.ResultsFertilization occurred in 138(69%) of 200 oocytes which were allocated to the chemical activation treatment groups. The activation rates of the oocytes in the ET, ET+6-DMAP, IO, IO+6-DMAP, 6-DMAP groups were 65.6%, 75.6%, 83.7%, 88.2% and 6.9%, respectively, showing that the activation rates of combining activation treatment groups were significantly higher than those of single activation treatment groups. Except the 1PN formation rate show a significantly difference with pronuclei formation between IO and IO+6-DMAP group, there was no other significantly differences with pronuclei formation among the groups. Sex chromosome analyses indicated 12 embryos with XX and 10 embryos with XY in 35 embryos derived from activated oocytes containing 2PN.Conclusions(1) It is possible to activate unfertilized oocytes after ICSI with chemicals, and the normal fertilized oocytes have further developmental potential; (2)ionomycin + 6-DMAP group showed the best activation rate among the groups, however we did not find any difference in developmental potential between the activation groups.Part II Study on the application of Oocytes Activation in Recue intracytoplasmic sperm injectionObjectiveTo investigate the application effect of Oocytes Activation in Rescue ICSI.DesignOocytes allocated to this study were those that did not show any signs of normal fertilization (extrusion of the second polar body, pronuclei formation) by 18h after IVF, and that were still with no signs of degeneration or fragmentation. 148 oocytes were randomly subjected to the study. The experimental group was that has total 92 oocytes which received chemical activation treatment after rescue ICSI , the control group has total number of 56 oocytes, without receiving chemical activation treatment after rescue ICSI. Fluorescence in situ hybridization (FISH) was employed to analyze chromosomes using X and Y dual-colored probes on the embryos that displayed two pronuclei in the experimental group.ResultsThe Fertilization rates in the experimental group and the control group were 89.1%(82/92)and 78.6%(44/56),the result showed that the fertilization rate of combining activation treatment after rescue ICSI was higher than the control group. The normal fertilization rate of the two groups were 51.2%(42/82)and 29.5%(13/44)respectively, which also had a significantly difference between the experimental group and the control group. Based on the embryos formed from normal fertilized oocytes of the two groups, no significantly difference were found in the cleavage rates and blastocyst rates. Sex chromosome analysis indicated 5 embryos with XX and 4 embryos with XY in 12 embryos derived from activated oocytes containing 2PN after rescue ICSI.ConclusionsThe combination of chemical oocytes activation treatment after rescue ICSI could effectively improve the fertilized rate after Rescue ICSI. and the normal fertilized oocytes have further developmental potential. However, we did not find any difference in developmental potential in the embryos formed from normal fertilized oocytes between two groups, showing that the oocytes activation can be applicated in unfertilized oocytes which receive rescue ICSI treatment. |
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