| Objective:To investigate the maturation, normal fertilization, abnormalfertilization, cleavage and high quality embryo rates of immature human oocytesderived from controlled ovarian hyperstimulation (COH) and intracytoplasmic sperminjection (ICSI) therapy cycles in vitro maturation (IVM) in control groupe anddifferent Activin-A(ACT-A) concentration (1 ng/ml, 10 ng/ml, 100 ng/ml) culturemedium. Analyse the best concentration and best culture time of ACT-A in IVM,furthermore finding out the mechanism action of ACT-A, providing experimentfundament for clinical; To observate and compare the ultrastructure of oocytes whichmatured in different concentration-time culture conditions, high quality and lowquality oocytes in different culture stages by transmission electron microscope(TEM). Criterion the maturation of oocyte membrane,periplast and nuclear, providingmorphology base for ACT-A in IVM and early embryo development.Method:1.78 IVF/ICSI-ET patients' 234 immuature oocytes were enrolled in thestudy; 2. According the different concentration of ACT-A (control group 0 ng/ml, 1ng/ml, 10 ng/ml, 100 ng/ml) and different culture time (24h, 30h, 48h), the objectsare devided into 12 groups randomly. In vitro maturation separately and record thematuration, normal fertilization, abnormal fertilization, cleavage and highquality embryo rates for statistics analysis using SPSS 11.5; 3. Observate theultrastructure of oocytes in differert experiment groups by TEM.Result:1. Culture time 30h group, beyond noamal fertilization rate has no statisticsdifferent between groups, for the rest all have statistics different between groups(P＜0.05). In groups, maturation rate of 100 ng/ml group significant higher thancontrol group (P＜0.005); normal fertilization rate of 100 ng/ml group higher than control group and 10 ng/ml group, having statistics difference (P＜0.05).Abnormal fertilization rate of control group significant higher than the rest groups,having significant statistics difference (P＜0.005); Cleavage and high qualityembryo rates of 100 ng/ml higher than control group and l ng/ml group, havingstatistics difference(P＜0.05). 2. Following the increasing concentration of ACT-A,the maturation, normal fertilization, cleavage and high quality embryo rates ofdifferent timing groups are advanced, and at the same time, abnormal fertilizationrates are depressed. The 30h group is the most significant. 3. Cleavage rate of ACT-A100 ng/ml group in different culture time has statistics different between groups(P＜0.05). Cleavage rate of 30h significant higher than 24h and 48h in group,having significant difference (P＜0.005); At the same time, high quality embryorate of 24h lower than 30h in group, having satistics difference(P＜0.05).Increasingculture time to 48h has no significant help to IVM and development of early embryo.4. All the observations seem to be have no concentration-time interact. There is nostatistics different(P＞0.05).5. Control group oocytes culturing for 24h show thatnuclear and periplast have significant derangement by TEM. The oocytes cultured in100 ng/ml ACT-A for 30h and 48h have better synchronism.Conclusion: 1. ACT-A has promotion effect in IVM and development of early embryo.Following the increase of ACT-A concentration, this effect is enhanced, showingconcentration dependent. 2. ACT-A 100 ng/ml culturing for 30h obtains satisfatiedoutcome. 3. Although prolong culture time can help the derangement of oocytesmaturation, but has no significant effect in maturation, normal fertilization rateand development of early embryo. Therefore, to be convenient in clinical, adopingculture time 30h is more scientific. 4. The immature oocytes derived from COH, ifcultured in suitable medium can also get satisfatied outcome. Even if there isdifference between the oocytes matured in vivo, but it increases the ET cycles anddecreases the cancle cycles. It has the energetic supplymental effect for IVF.
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