Marc Jg Van Den Bergh

The long-acting gonadotropin-releasing hormone analogues impaired the implantation rate.

OBJECTIVES To determine the efficacy and innocuousness of long-acting versus short-acting GnRH analogues (GnRH-a) in long protocol for in IVF-ET. DESIGN Prospective randomized study. SETTING The IVF unit at an academic hospital. PATIENTS One hundred couples admitted for their first IVF-ET attempt. MAIN OUTCOME MEASURES Serum concentrations of LH, E2, and P during the all cycles and duration of pituitary desensitization were assessed, as well as fertilization rate, embryo quality, and implantation and pregnancy rates. RESULTS Significantly more days (10.8 +/- 1.8 versus 9.2 +/- 1.7 days) of stimulation and more ampules of hMG (47 +/- 22 versus 33 +/- 16) were necessary to obtain similar numbers of embryos of quality with the long-acting GnRH-a. Implantation and delivery rates were significantly lower with the long-acting GnRH-a (32.8% versus 21.1%; 48.9% versus 29.1%, respectively). CONCLUSIONS As the long-acting GnRH-a might interfere with the luteal phase and embryo development, short-acting GnRH-a should be preferred for ovarian hyperstimulation in IVF-ET.

Incidence of Chromosomal Mosaicism in Human Embryos at Different Developmental Stages Analyzed by Fluorescence In Situ Hybridization

Chromosomal mosaicism has been reported in in vitro-cultured embryos at early cleavage stages, as well as in morulae and blastocysts. We have assessed the incidence and pattern of mosaicism during in vitro development of human embryos from early-cleavage stages to morula and blastocyst. Fifty spare embryos were fixed for fluorescence in situ hybridization (FISH) analysis for chromosomes X, Y, 13, 18, and 21 on days 2 or 3 (4- to 10-cell stage) (n = 16), on day 4 (morula stage) (n = 14), on day 5 (pre-expanded blastocyst) (n = 5), and the expanded blastocyst stages (n = 15). Blocked embryos (no cleavage observed within the last 24 hr) were not included. A total of 2367 cells were analyzed. Four early-cleavage stage embryos were found uniformly diploid; all of the others were mosaic for the chromosomes analyzed (mean diploid nuclei 48.3% +/- 28.7). All of the embryos at more advanced developmental stages, except one fully normal morula, had mosaic chromosome constitutions, with an increase in the percentage of diploid cells in morulae, pre-expanded, and expanded blastocysts, respectively (mean diploid nuclei 78.6% +/- 11.7, 66.0% +/- 20.8, 79.6% +/- 12.8), in comparison with earlier stages. Hypotheses about the origin of mosaicism and embryo regulation mechanisms will be discussed.

Noninvasive assessment of glucose and pyruvate uptake by human embryos after intracytoplasmic sperm injection and during the formation of pronuclei

OBJECTIVE To improve in vitro culture conditions and human embryo selection before transfer after IVF with intracytoplasmic sperm injection (ICSI). DESIGN A controlled, randomized, prospective study. SETTING University hospital-based IVF-ET program. PATIENT(S) Couples undergoing ICSI. INTERVENTION(S) Culture of human embryos in the presence of 1 mM or 5.56 mM glucose and metabolic measurements with the use of noninvasive microfluorescence assays immediately after ICSI to the time of transfer. MAIN OUTCOME MEASURE(S) Embryo development, implantation rate, and glucose and pyruvate uptake. RESULT(S) Fertilization rates, early embryo development, and implantation rates were not significantly different between 1 mM and 5.56 mM glucose. Pyruvate uptake was significantly higher during the formation of the pronuclei, at 15 +/- 0.7 and 11.4 +/- 1.3 pmol/embryo/h for fertilized and unfertilized oocytes, respectively. Pyruvate uptake did not correlate with cleavage stage or embryo morphology. However, during the second day of incubation, pyruvate uptake was significantly higher for the untransferred embryos of pregnant women compared with nonpregnant women, at 17.9 +/- 1.5 and 10.8 +/- 1.0 pmol/embryo/h, respectively. CONCLUSION(S) The increased level of pyruvate uptake during fertilization reflects the increased demand for energy necessary for the formation of the pronuclei. However, the metabolic measurements could not improve the selection of embryos with the best implantation potential. Finally, the reduction of glucose concentration in the culture medium failed to improve embryo viability.

Re-analysis by fluorescence in situ hybridisation of spare embryos cultured until Day 5 after preimplantation genetic diagnosis for a 47, XYY infertile patient demonstrates a high incidence of diploid mosaic embryos : a case report

Mosaicism in 4–8‐cell human embryos analysed by fluorescence in situ hybridisation (FISH) has been widely reported, but few studies have addressed the incidence of mosaicism in more advanced embryonic stages. In the present study we analysed spare human embryos in a case of preimplantation genetic diagnosis (PGD) for increased risk of aneuploidy because of an infertile 47,XYY man. After replacement of two embryos typed as 1818XX at PGD, six spare embryos (not frozen because of their low quality) were re‐analysed on Day 5 for PGD confirmation. Out of five embryos typed as 1818XY at PGD, four were diploid mosaic (DM) and one was normal in all cells. The sixth embryo, typed as 18XYY/1818181818X at PGD, was a DM. In spite of the bias of our small series of morphologically low‐quality embryos, the surprisingly high proportion of mosaics (which confirms previous findings) questions the validity of PGD, but supports the strategy of transferring only the embryos where two blastomeres gave normal and concordant results at PGD. More data are required to understand the clinical significance of early diploid mosaicism (and its impact on implantation rate) and to determine whether some diploid mosaic embryos might be considered safe for transfer. Copyright

Abnormal sperm-mucus penetration test predicts low in vitro fertilization ability of apparently normal semen

OBJECTIVE To investigate whether Kremers sperm-mucus penetration test may predict sperm fertilizing ability in IVF. DESIGN Kremers test was prospectively performed on semen samples used for 66 consecutive IVF trials and compared with the fertilization rates and fertilization failure rates observed. RESULTS Fertilization rates were significantly reduced in cases of abnormal Kremers test (42% versus 51%; n = 745 oocytes with a statistically insignificant increase in fertilization failure rates (21% versus 10%; n = 66 trials). For abnormal semen, fertilization rates (39% versus 39%; n = 208 oocytes) and fertilization failure rates (20% versus 28%; n = 17 trials) were similar regardless of Kremers test result. For normal semen, an abnormal Kremers test implied a significant decrease in fertilization rates (44% versus 54%; n = 537 oocytes) with a statistically insignificant increase in fertilization failure rates (21% versus 6%; n = 49 trials). CONCLUSIONS Abnormal Kremers test results identify patients with a decreased in vitro fertilizing ability despite apparently normal semen samples and a group with very low fertilizing failure risk in case of normal semen samples and normal Kremers test. Kremers test does not add any predictive value to sperm analysis in the case of abnormal semen samples. These observations point out the importance of the male factor in fertilization failure even in the case of normal semen analysis.

Second polar body extrusion is highly predictive for oocyte fertilization as soon as 3 hr after intracytoplasmic sperm injection (ICSI)

ObjectiveOur objective was to evaluate the time course and the predictive value of the extrusion of the second polar body after intracytoplasmic injection (ICSI) related to the fertilization rate, embryo cleavage and quality.SettingThe setting was the in vitro fertilization program of a university hospital.PatientsTwenty-one patients were treated with intracytoplasmic single sperm injection either for fertilization failure in IVF, low fertilization in IVF (<5%), or severe male factors.DesignOne hundred thirty-five of 205 metaphase 2 oocytes treated with intracytoplasmic single sperm injection were observed 1, 2, and 3 hr after the assisted fertilization procedure. Extrusion of the second polar body was recorded. For each of these oocytes, fertilization was noted 18 hr after ICSI and cleavage and embryo quality were assessed 24 hr later. The 70 remaining oocytes were used to assess a possible negative effect of repeated exposure to light microscopy.ResultsThe extrusion of the second polar body 3 hr after injection was an observation with a sensitivity of 0.87, a specificity of 0.58, and a high positive predictive value (0.90) toward oocyte fertilization. Twenty-nine and four-tenths percent of the oocytes extruded a second polar body within the first hour, 56.6% within the first 2 hr, and 78.3% had a second polar body 3 hr after injection. This time course was related neither to the speed of embryo cleavage nor to the embryo quality. Fertilization, cleavage, and embryo quality were not affected by repeated observation as deduced from comparison with the control group and confirmed by a high pregnancy (62% per oocyte retrieval) and implantation rate (22% per replaced embryo).ConclusionOocytes can be checked, in all safety, 3 hr after a single sperm injection for the presence of a second polar to predict oocyte fertilization with a high certainty.

Higher degree of chromosome mosaicism in preimplantation embryos from carriers of robertsonian translocation t(13;14) in comparison with embryos from karyotypically normal IVF patients.

AbstractPurpose: To compare the frequency and the degree of mosaicism in human embryos from Robertsonian translocation (RT) t(13;14) carriers, with embryos from karyotypically normal IVF patients. Methods: FISH analysis of embryos from PGD cycles for RT t(13;14), with probes for chromosomes 13, 14, and 18 (Group I) and of embryos from karyotypically normal IVF patients with probes for chromosomes 13, 18, 21, X, and Y (Group II). Results: The incidence of abnormal mosaic embryos was significantly higher in group I (38/51) as compared with group II (6/45) (χ2: P < 0.01). Furthermore, in group I the percentage of diploid cells per embryo was lower for chromosome 13 and 14 in comparison with 18, while in group II no differences were observed between the five chromosomes analyzed. Conclusions: RT induces a high frequency of mosaicism specifically for the chromosomes implicated in the translocation; the analysis by FISH of two blastomeres is strongly recommended for these patients.

The outcome of cryopreserved human embryos after intracytoplasmic sperm injection and traditional IVF

Objective:Our objective was to analyze the outcome of cryopreserved embryos obtained after intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) in terms of survival rate, implantation rate (IR), total and clinical pregnancy rate (PR) in a retrospective, comparative study.Methods:Three hundred seventy-five IVF and 463 ICSI surnumerary cleaved embryos, frozen on Day 2 with 1,2-propanediol, were thawed.Results:Thirty-two percent of the thawed IVF embryos survived and 11 pregnancies (8 clinical) were obtained from 68 transfers (16.1%). Fourty-seven percent of the ICSI embryos survived, with 19 pregnancies (18 clinical) from 116 transfers (16.4%). The IR was 8.5% (8/94) in IVF cycles and 10.8% (20/185) in ICSI cycles.Conclusions:A significantly better survival rate of ICSI embryos was observed but with no difference in PR, preclinical, and clinical abortion rate, or IR.

The Use of an Hydrogen Peroxide Multipurpose Isolator for Inhouse Preparation of Human Embryo Culture Media: A Unique Successful Swiss Randomized Prospective Study

Purpose: To develop inhouse made (IHM) embryo culture medium with a Multipurpose Isolator and compare the embryo development in a prospective randomized study with commercial media.Methods: Fertilization by intracytoplasmic single sperm injection (ICSI) of Metaphase II oocytes obtained after 96 controlled ovarian hyperstimulation cycles in patients not older than 37 years. Transfer of zygotes to IHM or commercial Cook Sydney IVF Cleavage medium (SIC) immediately after pronucleus observation. Evaluation of embryo cleavage and score, pregnancy, and implantation rate.Results: From 100 zygotes cultured in SIC, 61% were at the 4 cell stage 45 h after ICSI compared to 77% (78/101) in the IHM, P<0.05. The mean embryo score with IHM was 3.9±0.9 compared to 3.5±1.2 with SIC, P<0.05. The clinical pregnancy rate per transfer was 38.9% (37/95), the implantation rate was 23% (46/200), and no differences were observed between the groups.

PP-19 EVALUATION OF SPERM QUALITY AT A NATIONAL LEVEL: LOGISTIC, RECRUITMENT, ANALYTICAL AND STATISTICAL PROBLEMS ENCOUNTERED IN SWITZERLAND

PP-19 EVALUATION OF SPERM QUALITY AT A NATIONAL LEVEL: LOGISTIC, RECRUITMENT, ANALYTICAL AND STATISTICAL PROBLEMS ENCOUNTERED IN SWITZERLAND Josefina Vargas1, Roumen Parapanov1, Marc Van Den Bergh2, Eric Stettler3, Pierre Crettaz4, Alfred Senn1, Marc Germond1. 1Fondation Faber, Lausanne, Switzerland, 2Kantonspital Baden Ag, Fertilitatslabor, Switzerland, 3Swiss Army Medical Services, Switzerland, 4Federal Office of Public Health, Switzerland