| In vitro infertilization-embryo transfer(IVF-ET) development up to now, there are many new breakthroughs in COH ( controlled ovarian hyperstimulation) , the embryo culture technique and improvement of the culture medium , but how to increase the lower embryo implantation rate, clinical pregnancy rate, and reduce multiple pregnancy rate to perplex to reproduction medicine worker always.The blastocyst culture and transfer is a big progress of the under medical treatment in IVF-ET in near period . On abroad many the center of IVFs have been used as this the routine technique proceed the application. The local recent years also have opened the exhibition this research in a handful of laboratory.To find out implatation rate ,prenancy rate and multiple-gestation pregnancy rate of blastocyst transferred ,We adopted sequential culture system method in this study,30 cases were performed blastocyst transferred and 30 cases were performed cleavage stage embryos transferred. Two control group were studied with pair-matching.Material and MethodPatientsBetween September 2002 and February 2003, 30 women were selected randomly from patients who performed IVF-ET in our center , whose aged 25-35 (the average age 29.9 2.9) ,oocyte 10(average 13.8 3.8) , whose embryo transfers at the blastocyst stage were performed, defined as protocol group. 30 women whose embryo transfers at cleavage stage were control group. Use the method that form couples the design . The experiment set is in accordance with the matchedcontrol press the age differ 2 years old or soly, the MII period oocyte differ 21y or soly, COH project the similar proceeding in cause of infertility forms couples.Experiment methodControlled ovarian hyperstimulation was undertaken using subcutaneous buserelin acetate in a long protocol combined with Gonal-F. When the leading follicle reached 18mm in mean diameter with a serum estradiol level pf 1835mmol/ml,endomtrium thickness above 9mm ,10000IUof hCG was administered. Oocyte retrieval was performed 36 hours after the injection of hCG bytransvaginal ultrasonography. The sperm sample was washed with the Sper-mGrad by density gradient centrifugation, For the procedure, oocytes were inseminated with 5 (105/ml motile spermatozoa in IVF-20 medium. The matched control adopts routine culture method by using 1026 culture medium. All embryo transfers were performd 3 days after oocyte retrieval and embryo growth to 6-8 cell . Transferred 3 embryos. Blastcyst set adopt microdrop culture method by sequential media systems in medium 1Q26 and 1029 . Blastocyst quality was assessed in morning of 5 days after oocyte retrieval and 1-2 blastocyst embryos (the average 1.3 0. 6) was transferred . The luteal phase was supported with Progest-eron in two sets . Pregenancy was defined as a positive HCG test in urine on day 14 post transfer after embryo transfer . Clinical pregnancy was defined as the presence of a gestational sac ( s) with a viable embryo shown on vaginal ultrasonography performed approximately 28 days after embryo transfer. All patients were followed to 10 weeks in gestation.Experiment materialReagent:Quinn's Advantage?sequential media system 1023,1020,1026, 1029, HAS, SpermGrad?and deionized water were provided by the SAGE Bio-pharma company of the United States. Instrument: The German ZEISS Axio-vert200 dissecting microscop, the ZEISS Stemi2000 invert microscope, the MODEL3131 three air incubator of the United States ( C02, 02, N2) , MODEL3111 CO2 incubator, All culture dishs and test tubes were provided by the United States Becton Dickinson company.Data processing; All data and fixed amount datases input the computer, making use of the SPSS- PC statisticses the software proceeds statistics treatment , examining result by mean of the X ?S. The comparison of mean age, number of oocytes etc calculating marker were analyzed using T test and the comparison of rate were analyzed using X2 test between blastocyst set and control set.ResultsDifference was found in the implatation rates betwe...
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