De Grandi, P.

Delacretaz, G.

Descloux, L.

Germond, M.

Primi, M.-P

Rink, K.

Senn, A.

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Abstracts of the 15th Annual meeting of the ESHRE, Tours, France 1999Conclusion: ICSI outcome and pregnancy results are compar-able to those of IVF in patients with pure tubo-peritonealfactor infertility.to strict criteria were normal in all couples. All patientshad luteal-long gonadotrophin-releasing hormone analogueprotocol and standard procedures for ICSI and IVF. Chi-squareand independent t-tests were used for statistical analyses.Results: These are given in Table I.10.45-11.000-006. Computerized fast-motion video capture of mouseembryo development and hatching: a comparativeanalysis between normal and assisted hatchingPrimi M._P. I , Senn A.I , Rink K.2, Descloux L.3,Delacretaz G.3, De Grandi P.l and Germond M. I1Reproductive Medicine Unit, Dept. Obstet.-Gynaecol.,CHUV, CH-1011 Lausanne, 2Medical Technologies MontreuxSA, CH-1815 Clarens and 3Institut d'Optique Appliquee,EPFL, CH-1015 Ecublens, SwitzerlandIntroduction: Embryo development and hatching are difficultto observe as they take place inside an incubator and lastseveral days. Our goal was to develop a computerized videocapture system which allows the observation, recording andanalysis of these phenomena using fast-motion movies, whilekeeping the embryo in an environment (temperature, light,COb medium) suitable for its growth. Our first recordingswere aimed at comparing the development of control and zonapellucida (ZP)-drilled mouse embryos.Materials and methods: Zygotes are collected from femalemice and cultured for 1 day in carbonated human tubal fluidmedium supplemented with 5 mg/ml bovine serum albumin.ZP microdrilling is performed with a single 30 ms shot usinga 1.48 urn diode laser system (Fertilase; Medical TechnologiesMontreux, Switzerland), according to a technique describedelsewhere [Germond M. et ale (1995) Fertil. Steril., 64, 604-3OHSS on the DET. Pts with ~12 oocytes and/or oestradiol~2500 pg/ml were defined as high-risk-patients and separatelyrandomized to receive either protocols II or III. These groupswere defined as groups IV and V respectively. Complete dataof 241 pts were available at the time, when this abstractwas written.Results: 50, 42, 42, 48, and 59 pts were randomized toprotocol I, II, III, IV, and V, respectively. Pregnancy rates (PR)were 26.0, 19.0, 21.4, 27.1 and 28.8% in the five protocols.Overall PR was 24.9%, PRs in the low-risk (I-III) andhigh-risk groups (IV,V) were 22.4 and 28.0% respectively.Implantation rates (IR) were 10.6, 11.9, 8.8, 13.7 and 11.3%respectively. There was no significant difference between thegroups regarding PR or IR. Mean age was 31.84 ± 3.3,34.04 ± 3.8, 33.52 ± 4.2, 31.02 ± 3.3, and 31.39 ± 4.0years, respectively. There was a significant difference (Kruskal-Wallis test, KWT) between group II versus groups IV and V,as well as between groups III versus IV (P < 0.05). Bodymass index was not different between the groups. Meanoestradiol was significantly higher in groups IV and V, com-pared to groups I, II, and III (P < 0.05). The same held forthe number of oocytes retrieved (7.40 ± 2.6, 6.71 ± 3.2,7.55 ± 2.9, 14.78 ± 5.2, 14.59 ± 5.3, in groups I, II, III,IV, and V respectively). The cumulative embryo score wassignificantly higher in group V (29.95 ± 8.3), compared togroup 11 (22.31 ± 11.7) and III (23.33 ± 10.5) (KWT, P <0.05). Two severe OHSS were observed, one each in groupsIV and V. Three moderate OHSS occurred in the low-riskgroup, all in group I, and one occurred in group V. Nineteenmild OHSS were recorded, two in each of the low-risk groups,six in group IV and seven in group V.Conclusion: There is no indication to use HCG in LPS, sincethere is no significant difference in PR between the groups.There is a higher risk of OHSS after HCG administration. Theslight, non-significant differences observed between the PRare mainly due to the differences in age and embryo quality,which occurred despite a randomization system.10.30-10.450-005. Intracytoplasmic sperm injection versus in-vitrofertilization in pure tubal factor infertility: a prospectiverandomized trialBukulmez 0., Yucel A., Yarali H., SarI T., Girgin B.,Gunalp S. and Gurgan T.Hacettepe University, Faculty of Medicine, Department ofObstetrics and Gynecology, Slhhiye 06100, Ankara, TurkeyIntroduction: It is highly controversial whether intracyto-plasmic sperm injection (ICSI) will replace standard in-vitrofertilization (IVF) in patients with pure tubal factor infertility.The aim of this prospective randomized study is to compareICSI versus IVF in women with tubo-peritoneal factor infer-tility.Materials and methods: Patients with tubo-peritoneal factorinfertility were randomized to ICSI (n = 38) or IVF (n = 38).Semen analysis including morphology assessment accordingTable I. Baseline characteristics and ICSIIIVF results.ICSI-ETEmbryo transfer cycles (n) 38Female age (years) 34 ± 3.96Male age (years) 37.46 ± 4.15Duration of infertility (years) 10.08 ± 4.19Patients with unilbilateral hydrosalpinx (n) 4.0Oestradiollevel-HCG (pmol/l) 5193.7 ± 3764Oocytes (n) 9.08 ± 6.22Metaphase-II oocytes (n) 7.32 ± 4.342-pronuclei rate (%) 69.03 ± 22.12Cleavage rate (%) 66.87 ± 23.04Embryos (n) 4.97 ± 3.06Grade-I embryos (n) 3.45 ± 2.43Embryos transferred (n) 3.39 ± 1.39Grade-I embryos transferred (n) 2.71 ± 1.71Individual implantation rate (%) 38.75 ± 24.46Clinical pregnancy/ET(%) 21.05Take-home babylET (%) 18.42ET =. embryo transferAll results are non-significant.IVF-ET3834.03 ± 3.7737.34 ± 4.759.06 ± 4.594.05071 ± 30608.11 ± 3.77.0 ± 3.567.32 ± 25.0169.84 ± 23.735.0 ± 3.173.13 ± 2.543.61 ± 1.592.5 ± 1.4334.58 ± 16.9721.0515.79Abstracts of the 15th Annual Meeting of the ESHRE, Tours, France 1999611]. A single 2-cell embryo is transferred into aID ul culturemedium droplet under mineral oil inside a Petri dish, fittinginto an in-house constructed humidified micro-incubator, main-tained at 37°C by water circulation and gassed with 5% CO2.A CCD video camera mounted on the microscope sends asignal to a frame grabber card located in a computer. Theimage is digitized with a time lapse of several minutes andstored as a file with a codified name generated automaticallyby the computer software. Microscope light source and imageacquisition frequency are controlled by visual basic software(PA Images, Swltzerland), which also generates the moviesfrom the stored image sequences.Results: The specially designed micro-incubator offers anenvironment compatible with embryo development. Normalcleavage rates and hatching occur over the 4-5 days of culturewith only a need for medium renewal. In control embryos,several extension/compaction movements (ECM) led to aprogressive thinning and to a sudden large slitting of the ZPthrough which the expanded blastocyst quickly escapes. Thelaser-drilled embryos start hatching 1 day earlier than thecontrols by protruding through the drilled hole. The ECM donot lead to ZP thinning and rupture; more ECM are neededbefore hatching is completed.Conclusion: We hereby present a new research tool whichallows us to study the hatching phenomenon as well as all theother stages of the in-vitro development in embryos of allspecies. The computerized images provide easily accessiblematerial for different measurements. The movies can be madeover longer periods of time than normal videos as they arecompiled to accelerate phenomena usually difficult to observe.11.00-11.150-007. Maturation in-vitro of mouse prenatal folliclesrecovered from ovarian grafts under the kidney capsuleLiu J., Van der Elst J., Van den Broecke R., Dumortier F.and Dhont M.Infertility Centre, Department of Obstetrics and Gynecology,Ghent University Hospital, De Pintelaan 185, 9000 Ghent,BelgiumIntroduction: Thousands -of primordial follicles are presentin the ovaries of neonatal mammals. Until now, reproducibleconditions for a complete development of primordial folliclesin vitro have not been established. The aim of this study wasto achieve complete development of primordial follicles innewborn mouse ovaries by combining in-vitro and in-vivosteps. A two-step strategy was set-up. First the ovaries fromnewborn mice were transplanted heterotopically under thekidney capsule of ovariectomized adult female recipients; after14 days, preantral follicles were isolated mechanically fromthe recovered grafts and cultured in vitro for 12 days.Materials and methods: Six- to eight-week-old F1 (C57BL/6jXCBAlCa) female ovariectomized mice were used as recipi-ents. Newborn F1 female mice (n = 6) were killed andthe ovaries were dissected free. Five ovaries that were nottransplanted were placed into Bouin's fluid and processed for4histological examination. One newborn mouse ovary wastransplanted under the renal capsule of each recipient (n =7). Two weeks after transplantation, preantral follicles wereisolated from four ovarian grafts and three recovered graftovaries were prepared for histological examination. The pre-antral follicles were cultured individually in 20 ul a-MEMmedium supplemented with 100 mIU/ml recombinant follicularstimulating hormone (rFSH) and 5% FCS in 60 mm petridishes incubated at a condition of 37°C, 100% humidity, and5% CO2 in air for 12 days. Half of the medium was refreshedevery 2 days. At day 12 of culture, final oocyte maturation wasinduced by adding 2.5 IU/ml human chorionic gonadotrophin(HCG). The nuclear maturation of oocytes was assessed afterdenudation from granulosa cells. Preantral follicles (n = 97)isolated from 14-day-old mice were cultured under the sameconditions as control group. X2 test was used for statisticalanalysis.Results: All grafts (n = 7) were recovered 2 weeks aftertransplantation. Histological examination of recovered grafts(n = 3) revealed that follicles proceeded to preantral andantral follicle stages with a predominance of preantral follicles.Approximately 15:-25 preantral follicles were isolated mechan-ically from each ovarian graft (n = 4) and total 64 selectedpreantral follicles (100-150 urn in diameter) were put intoculture. There were differences between the control and thetransplanted groups in the percentage of follicle survival during12 days in-vitro culture (98 versus 64% repectively; P <0.0001), and in the percentage of oocytes that acquiredcompetence to undergo germinal vesicle breakdown (GVB)(91 versus 65% respectively; P < 0.0001) or in the percentageof GVB oocytes that produced a polar body, indicating progres-sion of meiosis to metaphase 11 (86 versus 46% respectively;P < 0.001).Conclusion: The results reported here show that maturation ofmouse primordial follicles is possible by in-vivo transplantationcombined with in-vitro culture. This two-step method may bea potentially attractive alternative to promote the transitionfrom primordial to preantral follicles that can be supported byan in-vitro culture system.11.15-11.300-008. Impact of stages III-IV of endometriosis on theoutcome in recipients of oocytes from the same donor:matched case-control studyDiaz 1.1, Navarro J.1, Simon C. 1,2, Pellicer A.l,2 andRemohi J.1,2'Lnstituto Valenciano de Infertilidad (IVI), and 2Departmentof Paediatrics, Obstetrics and Gynaecology, ValenciaUniversity School of Medicine, Valencia, SpainIntroduction: Infertility in endometriosis patients has beenattributed to alterations within the oocyte which, in tum, resultin embryos of lower quality and reduced ability to implant.The aim of this study was to compare the in-vitro fertilizationand embryo transfer (IVF/ET) outcome in oocyte recipientswith and without severe endometriosis receiving eggs fromthe same donor.

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