First singleton term birth after ovarian superovulation with rhFSH.
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Biomedical subjects
Publications and source records attributed to P Devroey.
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Intracytoplasmic sperm injection (ICSI) is a promising assisted-fertilisation technique that may benefit women who have not become pregnant by in-vitro fertilisation (IVF) or subzonal insemination (SUZI) of oocytes. We have used ICSI to treat couples with infertility because of severely impaired sperm characteristics, and in whom IVF and SUZI had failed. Direct injection of a single spermatozoon into the ooplasm was done in 47 metaphase-II oocytes: 38 oocytes remained intact after injection, 31 became fertilised, and 15 embryos were replaced in utero. Four pregnancies occurred after eight treatment cycles--two singleton and one twin pregnancy, and a preclinical abortion. Two healthy boys have been delivered from the singleton pregnancies and a healthy boy and girl from the twin pregnancy.
Diagnosis of genetic disorders in the embryo before implantation, though possible by removal of one or two blastomeres at the eight-cell stage, is still experimental because the procedures of gene analysis of DNA from a single cell are not yet reliable enough for clinical application. We have evaluated the efficiency and accuracy of polymerase-chain-reaction (PCR) amplification of a single-copy gene, wild-type or cystic fibrosis delta F508 allele, on single sperm cells from a donor known to be a heterozygous carrier of the delta F508 mutation. DNA from single spermatozoa was decontaminated by restriction-enzyme treatment, then the region around the delta F508 site was amplified by nested PCR. The distribution of the wild-type and mutant alleles (59 [55%] and 48 [45%, respectively]) in the 107 single spermatozoa did not differ from that expected (50% each). 1 sample did not provide an amplified signal. To check that the two alleles would be amplified with equal efficiency when they were both present within a cell, we did PCR for 51 two-sperm samples. Again the distribution did not deviate from that expected (17 [33%] both wild-type; 21 [41%] one wild-type, one delta F508; 13 [26%] both delta F508 vs 25%; 50%; 25% expected). None of the 74 blanks in these experiments was contaminated. We conclude that our delta F508 single-cell assay is efficient and accurate and can be used for analysis of blastomere DNA to diagnose cystic fibrosis before embryo implantation.
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OBJECTIVE: To compare zygote intrafallopian transfer (ZIFT) and in vitro fertilization and embryo transfer (IVF-ET) as treatments of male-factor infertility. DESIGN: Patients were prospectively randomized to ZIFT or IVF-ET. SETTING: In vitro fertilization program of the Centre for Reproductive Medicine of the Dutch-speaking Brussels Free University, Belgium, which is a tertiary referral institution. PATIENTS: One hundred fifty-seven couples were enrolled in the study. Inclusion criteria allowed only first trials of couples with long-standing infertility caused by a male factor. Female factors were excluded. INTERVENTIONS: In ZIFT, up to three fertilized oocytes were transferred into one single patient fallopian tube by means of laparoscopy 18 hours after insemination. In IVF-ET, cleaving embryos were replaced into the uterine cavity about 48 hours after insemination. MAIN OUTCOME MEASURES: Fertilization and transfer rates, implantation and pregnancy rates, pregnancy outcome, and cost per procedure were evaluated. RESULTS: Implantation rates of 12.3% and 10% per replaced conceptus were achieved for ZIFT and IVF-ET, respectively. CONCLUSIONS: This study demonstrates no therapeutic advantage of ZIFT over IVF-ET in male-factor infertility in terms of reproductive outcome or economic benefit.
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This chapter describes general cryobiological principles and the different methods for cryopreservation of supernumerary human embryos obtained after several procedures for medically assisted procreation. Different factors that influence the performance of freezing and thawing of human embryos are reviewed: the data from international and national surveys of results, the cryopreservation procedures, the stages of embryonic development, the morphological appearance of the cryopreserved embryos, the numbers of embryos transferred, the storage times of embryos in liquid nitrogen, the ovarian stimulation protocols in the IVF cycle and the replacement cycles of cryopreserved embryos. Cryopreservation of embryos circumvents the difficult problem of synchrony between the ovarian cycles of donor and acceptor patients in an oocyte donation programme.
Although healthy babies have been born after preimplantation diagnosis for sex determination, this technique is still in an experimental phase. To date, removal by micromanipulation of one or two blastomeres from an eight-cell embryo seems an acceptable procedure. Since a reasonable number of biopsied embryos will implant and develop in utero, the methodology for the analysis of the blastomeres at the chromosomal, gene product or DNA level, needs to be improved before further clinical application. In the meanwhile, more information about early embryo development will become available.
Laparoscopic removal of benign mature teratoma is safe and effective in pre-menopausal patients. Fourteen patients underwent laparoscopic removal of such tumours. There were no operative complications. Post-operative hospitalization was short (average: 2 days). All patients undergoing a second-look laparoscopy had mild adhesions around the operated ovary. Two spontaneous and one in-vitro fertilization pregnancy occurred. If a teratoma is diagnosed during laparoscopy or oocyte retrieval, it should be promptly removed as malignant transformation is not excluded. In this case, the embryos should be frozen for later transfer.
A luteal phase defect has been demonstrated in cycles stimulated using a protocol including a gonadotrophin releasing hormone agonist (GnRHa). We have conducted a randomized prospective study of luteal and early pregnancy supplementation in 262 women selected for in-vitro fertilization (IVF), gamete intra-Fallopian transfer (GIFT) or zygote intra-Fallopian transfer (ZIFT). Either intramuscular progesterone in oil (50 mg/day) or intravaginal micronized progesterone (600 mg/day) was used as luteal supplement. In association with oestradiol valerate, progesterone administration was initiated from the day before oocyte retrieval until the 12th week of pregnancy. The implantation rate just failed to reach statistical significance (P = 0.07) in favour of the group receiving intravaginal progesterone. In the latter group, we observed a higher clinical pregnancy rate (33.6 versus 26.7%, not significant). Despite lower plasma progesterone levels, a lower first trimester abortion rate (P less than 0.05) was found in the intravaginally treated group. Intravaginal micronized progesterone was well tolerated by all patients and appeared more effective than intramuscular progesterone in improving the implantation rate, and in decreasing the incidence of abortions in stimulated cycles including GnRHa.
The in-vitro fertilization and embryo transfer (IVF-ET) procedure was compared in 175 couples with male infertility and 480 couples with tubal infertility. In cases of male infertility, more oocytes were recovered but fewer oocytes were fertilized. Although the cleavage rate was decreased, no difference in embryo quality was found. In male infertility, fewer transfers were performed and the average number of embryos per transfer was lower. The total pregnancy rate was also lower per cycle (12.8 versus 22.9%), but not per transfer (25.4 versus 25.7%). The further obstetrical outcome was similar in both groups. We conclude that male infertility can be treated by IVF-ET but results are still disappointing when compared to a control group with normal spermatozoa.
Human spermatozoa were injected into the perivitelline space of oocytes from 43 couples (44 cycles) in whom fertilization had failed in conventional in-vitro fertilization (IVF). The spermatozoa were treated to enhance the percentage of acrosome-free spermatozoa either by incubation for 24 h in T6 medium with 50% follicular fluid (v/v) or by incubation for 24 h in T6 medium followed by electroporation and incubation for a few hours in T6 medium with 3.5 mM pentoxifylline. After these two procedures, the mean percentage of acrosome-free spermatozoa increased to 35.5 and 53.9% respectively. Up to three spermatozoa were injected into the perivitelline space of metaphase II oocytes; few oocytes were damaged during the injection procedure. The overall fertilization rate was 30.9% of the 433 oocytes that were intact after subzonal insemination. Only 3% of the injected oocytes had more than two pronuclei. The cleavage rate of the fertilized oocytes was 80%. There was no difference in the fertilization and cleavage rates between the two sperm treatment procedures. One, two or three embryos were replaced in 34 cycles and seven patients became pregnant. In three of the four ongoing pregnancies, prenatal diagnosis by amniocentesis indicated a normal karyotype.
Sixty-three normo-ovulatory infertile women were randomly divided into two groups. All women were first desensitized with the gonadotrophin-releasing hormone agonist (GnRHa), buserelin. Thereafter, ovarian stimulation with human menopausal gonadotrophins (HMG) was started in both groups but in group A the GnRHa was stopped on the same day. In group B, the GnRHa was continued during HMG treatment until the ovulatory human chorionic gonadotrophin stimulus was given. Premature luteinization was not observed in either group, although the preovulatory basal luteinizing hormone (LH) secretion was significantly higher in group A. An equal number of embryos of comparable quality was transferred in both groups and the pregnancy outcome was similar. However, the supernumerary embryos of group A were of a lower morphological quality and survived the cryopreservation process less well. We concluded that the continuous administration of a GnRH agonist during HMG treatment resulted in better quality of supernumerary embryos.
A total of 199 patients (412 consecutive cycles) were treated by oocyte-embryo donation in 336 replacement cycles. Of these, 296 involved intra-uterine embryo transfers, 38 zygote intra-Fallopian transfers (ZIFT) and two gamete intra-Fallopian transfers (GIFT). Of the 336 replacements, 244 (73%) constituted transfers of fresh concepti and 92 (27%) of frozen-thawed ones. A total of 85 pregnancies were achieved of which 16 ended in preclinical abortions, giving a clinical pregnancy rate of 34.7% per patient, 20.5% per transfer and a take-home baby rate of 29.1% per patient. The pregnancy rate was significantly higher (P less than 0.05) following fresh gamete or embryo replacement (23%; 56/244) than following that of frozen-thawed embryos (14.1%; 13/92). No significant difference was observed when intra-Fallopian replacement was applied (27.5%; 11/40) as opposed to intra-uterine (19.6%; 58/296). Ovarian function was not found to be of significant importance to the achievement of pregnancies after oocyte donation since comparable pregnancy rates per replacement and per started cycle were obtained in patients with ovarian failure and in those with functional ovaries (19% and 15.4%; 24.2% and 20.2% respectively). Comparison of the implantation and abortion rates between these two groups did not reveal any significant difference (11.1% and 11.1% versus 14.8% and 16.6%). The highest pregnancy rate among patients with ovarian failure was observed in those with primary ovarian failure (26.4%; 14/53), while the lowest was among women who had received chemotherapy and/or radiotherapy (9%; 1/11).(ABSTRACT TRUNCATED AT 250 WORDS)
Interceed (TC7) is a surgical adjuvant which has been shown to be efficacious in reducing adhesion formation in gynaecological pelvic surgery by laparotomy. In view of the increasing number of laparoscopic interventions and the concern with reducing post-operative adhesions, we evaluated the possibility of Interceed delivery through the ancillary trocars. Twenty-four patients underwent operative laparoscopy followed by the application of Interceed. No difficulties were noticed. Material sized from 1.9 x 2.5 to 3.8 x 5 cm was successfully delivered into the peritoneal cavity, while the duration of the procedure ranged from 2.5 to 6 min for each piece. In conclusion, it is feasible to consider application of Interceed during laparoscopic interventions. This approach to delivery requires a short operative time, minimal instrumentation and enhanced ease of application.
Pituitary gonadotrophin reserve and basal gonadotrophin secretion were tested during the luteal phase in women superovulated with buserelin/human menopausal gonadotrophin (HMG) in a desensitization (n = 17) or flare-up protocol (n = 7). In the desensitization protocol the luteinizing hormone-releasing hormone (LHRH) stimulated serum LH and follicle stimulating hormone (FSH) concentrations remained impaired at least until day 14 after arrest of the agonist. In the flare-up protocol basal and stimulated LH secretion was still abnormal on days 14 and 15 after human chorionic gonadotrophin (HCG) injection. Normal basal serum FSH concentrations were measured at the end of the luteal phase in the flare-up protocol, but the response of FSH to LHRH injection was still subnormal. We conclude that gonadotrophin function remained impaired until the end of the luteal phase after desensitization and flare-up GnRH-agonist and HMG stimulation protocols. Corpus luteum stimulation with exogenous HCG or substitution therapy using natural progesterone are required to prevent the possible negative effects resulting from pituitary dysfunction after GnRH-agonist treatment.
A successful zygote intra-Fallopian transfer has been performed using spermatozoa from a patient with Kallmann's syndrome. Spermatogenesis was induced by hormonal treatment but even so, semen quality remained slightly impaired and no pregnancy occurred over 4 years, despite ovulation induction in the patient's spouse. Finally she conceived after assisted procreation by in-vitro fertilization and delivered a healthy baby.
beta-N-acetylhexosaminidase is a lysosomal enzyme, which has two isoenzymes: beta-Hex A, a trimer consisting of one alpha-chain and two beta-chains (alpha beta 2) and beta-Hex B, a tetramer formed of four beta-chains (beta 2 beta 2). Genetic defects in the alpha-chain lead to Tay-Sachs disease, whereas mutations in the beta-chain gene lead to Sandhoff disease. In a previous study we developed a microassay for total beta-N-acetylhexosaminidase and used this for measuring activities in mouse oocytes and preimplantation embryos. In this study, to assess the feasibility of transferring this technique to the human for the purposes of preimplantation diagnosis for Tay-Sachs and Sandhoff disease, beta-Hex activity was assayed in human oocytes and embryos and in the medium in which they had been cultured. We showed that although the activity of beta-N-acetylhexosaminidase in human oocytes and embryos was > 500 times higher than in the mouse, it was not detectable in the culture medium and the activity in oocytes and embryos remained virtually constant throughout human preimplantation development, making it difficult to distinguish embryonic from maternal enzyme activity. In the absence of this distinction it would be inappropriate to use beta-N-acetylhexosaminidase activity for the purposes of preimplantation diagnosis of Sandhoff or Tay-Sachs disease. These experiments demonstrate that measuring the beta-N-acetylhexosaminidase activity in human embryos cannot be used at present for preimplantation diagnosis.