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[The use of clomiphene and menotropins as inducers of multiple follicular growth in a program of in vitro fertilization].

In our IVF & related techniques program, clomiphene 50 mg was used from days 2 to 6 of the cycle in 20 patients and 20 cycles. (17 IVF patients and 3 GIFT patients) menotropins were also used in 2 different schemes: a) 2 ampules in alternated days (12 cases) and b) one ampule daily (8 cases). The patients with the daily scheme required lesser total dose of menotropins and developed higher estradiol levels, although this had none statistic significance, and they also developed a greater number of follicles. There were 16 laparoscopies for ovum capture and 53 oocytes were retrieved (30.2% mature) 10 oocytes were from the GIFT patients. 16 oocytes were fertilized and 8 cleaved. There was only one pregnancy from and IVF patient with the alternated scheme that ended with a first trimester abortion.

Adult↗

A prospective randomized comparison of intramuscular or intravaginal natural progesterone as a luteal phase and early pregnancy supplement.

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.

Administration, Intravaginal↗

Factors affecting pregnancy rates of in vitro fertilization and gamete intrafallopian transfer.

Factors affecting pregnancy rates in gamete intrafallopian transfer (GIFT) and in vitro fertilization (IVF) were evaluated. Higher pregnancy rates were found when more mature oocytes were recovered (6.2 +/- 2.5 vs 3.8 +/- 3.0) and more oocytes (4.6 +/- 1.0 vs 3.3 +/- 1.2) or zygotes (4.7 +/- 1.4 vs 3.0 +/- 1.7) were transferred. Ultrasound-guided follicular aspiration was successful for in vitro fertilization and had a pregnancy rate (14.3%) comparable to the laparoscopic approach (13.3%). Laparotomy for correction of pelvic pathology concomitant with oocyte retrieval should be used cautiously despite the high preliminary pregnancy rate (42.9%). Failed fertilization of the surplus oocytes left after gamete intrafallopian transfer did not mean a poor chance of pregnancy (27.6%). The combined treatment of gamete intrafallopian transfer and in vitro fertilization may have a higher chance of pregnancy (43.5% vs 21.4% in GIFT and 13.3% in IVF), and more multiple pregnancies (40.0% vs 22.2% in GIFT and 16.7% in IVF). Cycles with spontaneous LH (luteinizing hormone) surges, detected by daily morning urine samplings, need not be aborted and the timing of ovum recovery should be individually evaluated.

Female↗

An improved medium for long-term culture of human embryos overcomes the in vitro developmental block and increases blastocyst formation.

OBJECTIVE: To determine if a culture medium, designated CZB after the authors who first described it, which is supplemented with 0.11 mM ethylenediaminetetraacetic acid, 1.0 mM glutamine, 31.30 mM lactate, and 0.27 mM pyruvate and is lacking glucose for the initial stages of culture that overcomes the in vitro two-cell block of mouse embryos, can improve the rate of blastocyst formation of human embryos in long-term cultures and increase the pregnancy rate (PR) when used in a clinical in vitro fertilization (IVF) program. DESIGN: The study is in two parts. Initially, excess oocytes from IVF and gamete intrafallopian transfer patients were fertilized in vitro and then placed in long-term culture of either CZB plus 10% heat-inactivated human serum (32 zygotes) or Earle's balanced salt solution (EBSS) supplemented with 0.45 mM pyruvate plus 10% human serum (28 zygotes) to determine if CZB medium could enhance in vitro development and increase blastocyst formation when compared with EBSS. Subsequently, CZB or EBSS medium was used for short-term cultures of embryos in a clinical IVF program to determine if the use of CZB could increase the clinical IVF PR. SETTING: Private practice of one author (M.D.). PATIENTS: The excess oocytes were donated by couples not wishing to have cryopreservation. In the clinical trial, 49 couples presenting with tubal or male factor infertility and who had three or more fertilized zygotes were randomly assigned to one of the culture media being used. RESULTS: In long-term cultures, embryos were observed at 42, 66, 90, 114, and 138 hours after fertilization and scored for blastomere number, degree of fragmentation, and developmental arrest. When CZB- and EBSS-cultured embryos were evaluated over 138 hours, there was a significant increase in the number of CZB-cultured embryos reaching the blastocyst stage (56% versus 20%; P less than 0.009) and less fragmentation of CZB-cultured embryos (18.8% versus 50%; P less than 0.01). In short-term cultures, pronuclear stage embryos from patients undergoing IVF were cultured in CZB or EBSS for 24 hours, graded, and then used in embryo transfers. Of the 28 patients assigned to EBSS, 6 became pregnant (21.4%), and of the 21 assigned to CZB, 5 attained pregnancy (23.8%). These results were not significantly different. CONCLUSIONS: The use of CZB for the long-term culture of human embryos is highly beneficial and increases the rate of blastocyst formation, but its use in an IVF program does not increase the clinical PR.

Analysis of Variance↗

Biological factors affecting subzonal sperm microinjection results.

One hundred and sixteen couples with severe male factor infertility underwent 139 subzonal sperm microinjection cycles. In total, 1343 oocytes were microinjected, resulting in a fertilization rate of 24%, followed by a cleavage rate of 65%. In 26% of the zygotes, fertilization was delayed and embryos derived from these zygotes demonstrated a poor capacity for further growth and implantation. In 102 of 139 cycles (73%) embryo transfer was performed, resulting in 9 pregnancies. This study followed the fate of injected oocytes and early embryo development to investigate biological factors that influence the results of subzonal injection.

Embryo Transfer↗

In-vitro fertilization, gamete- or zygote intra-fallopian transfer for the treatment of male infertility.

This retrospective study compares in-vitro fertilization and embryo transfer (IVF-ET), gamete intra-Fallopian (GIFT) and zygote intra-Fallopian transfer (ZIFT) for the treatment of male infertility in 266 couples (n = 318 cycles). The IVF-ET technique revealed a total pregnancy rate of 14.1% per cycle and 29.1% per transfer. The conception rate with GIFT was 17.8%; an abortion rate of 50% was noted. The ZIFT procedure scored a 20.9% total pregnancy rate per cycle and 33.3% per transfer. The cumulative take home baby rates per couple for IVF-ET, GIFT and ZIFT were 13.5, 7.0 and 20.0%, respectively.

Adult↗

Chromosome studies and fertility treatment in women with ovarian failure.

In vitro fertilization and embryo transfer or gamete (or zygote) intra-Fallopian transfer after ovum donation were performed in 16 patients with primary or secondary amenorrhea, associated with chromosome abnormalities. The patients showed the wide range of (mostly X) chromosome abnormalities characteristic for women with primary or premature ovarian failure. Four of these patients became pregnant and three of them have delivered healthy infants with a normal karyotype. This pregnancy rate is far superior to the accepted fertility figure in these patients. When these results were compared with the fertility treatment results of three other groups of women with absent ovarian function (1. ovarian dysgenesis; 2. surgical castration; 3. premature menopause) but with a normal 46,XX karyotype, no difference in treatment efficiency could be detected. These results offer a promising approach for the treatment of infertility in agonadal patients with chromosome aberrations.

Adult↗

The purpose of prelaparoscopy transvaginal retrieval and evaluation of oocytes in a modified GIFT procedure.

To minimize the risk of submitting the patient to an 'unnecessary' laparoscopy in cases of unsuccessful oocyte retrieval and to reduce gamete exposure to the probable adverse effects of the CO2 pneumoperitoneum and anaesthetic agents, we have performed transvaginal oocyte recovery prior to performing the laparoscopy for gamete intra-Fallopian transfer (GIFT). When the recovered oocytes had a low potential for fertilization, the GIFT laparoscopy was cancelled and fertilization in vitro and zygote intra-Fallopian transfer (ZIFT) were attempted instead. By introducing this modified, two-stage GIFT procedure, the laparoscopy time was reduced to a quarter of that required for the more time-consuming, conventional GIFT method (18 +/- 4 min and 76 +/- 13 min respectively in the same unit). Of the 35 cases selected for this modified GIFT treatment, only 26 were considered suitable to proceed to laparoscopy, resulting in eight clinical pregnancies (30.7%). The remaining nine (25.7%) GIFT cases were cancelled, due either to the absence of oocytes in four or to their poor quality in five (oopenia). The latter were diverted to IVF and out of five cases, only two oocytes were fertilized and proceeded to ZIFT. No clinical pregnancy resulted. In conclusion, we propose that securing the retrieval of oocytes of appropriate quality seems to be a logical and necessary preliminary step before embarking upon laparoscopy for GIFT or ZIFT, depending upon the biological status of the gametes. By employing this procedure, unnecessary and costly laparoscopy can be avoided in certain cases.

Carbon Dioxide↗

The stages at which human fertilization arrests: microtubule and chromosome configurations in inseminated oocytes which failed to complete fertilization and development in humans.

The goal of fertilization is the union of one, and only one, sperm nucleus with the female pronucleus within the activated oocyte. For this to occur successfully, several events must transpire, including the incorporation of the entire spermatozoon into the oocyte, the completion of meiotic maturation with the extrusion of the second polar body, the metabolic activation of the previously quiescent oocyte, the decondensation of the sperm nucleus and the maternal chromosomes into the male and female pronuclei respectively, and the cytoplasmic migrations of the pronuclei, which bring them into apposition. Defects in any of these events are lethal to the zygote and might prove to be causes of infertility. In this study, the microtubules and DNA were imaged in inseminated human oocytes that had been discarded as unfertilized. The presence and number of incorporated sperm tails were also documented using a monoclonal antibody specific for the post-translationally modified acetylated-alpha-tubulin found in the tail, but not the oocyte, microtubules. An analysis of 211 oocytes from failed in-vitro fertilizations from 58 patient couples resulted in the determination of several previously undetectable phases at which fertilization arrests: (i) metaphase II arrest; (ii) arrest after the successful incorporation of the spermatozoon, (iii) arrest after the formation of the sperm aster; (iv) arrest during mitotic cell cycle progression; and (v) arrest during meiotic cell cycle progression. Data on polyspermy and arrested embryonic development are also presented. These results have implications for the diagnosis and treatment of female, as well as male, infertility.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Cycle↗

Profiling assisted reproductive technology: outcomes and quality of infertility management.

OBJECTIVE: To critically appraise the content of the American Society for Reproductive Medicine (ASRM)/Society for Reproductive Technology (SART) Registry. DESIGN: English-language literature review. PATIENT(S): Women undergoing treatment with assisted reproductive technology (ART). INTERVENTION(S): Current ART treatments, including IVF, GIFT, zygote intrafollopian transfer (ZIFT), oocyte micromanipulation, and cryopreserved embryo transfers. MAIN OUTCOME MEASURE(S): Compliance with clinical practice guidelines, and casemix-adjusted rates of live delivery, clinical pregnancy, ectopic pregnancy, miscarriage, birth defects, implantation, fertilization, and retrieval. RESULT(S): Outcomes should be adjusted for variation in patient characteristics known to affect prognosis, including maternal age, the duration of infertility, the presumed cause(s) of infertility, the patient's prior history of treatment for infertility, and diethylstilbestrol exposure. Outcome rates should be reported using the patient as the denominator, as well as cycle, retrieval, and transfer. The statistical significance of observed differences in events rates should be indicated. Because widely accepted clinical practice guidelines related to performance of ART procedures are not available, compliance with practice guidelines cannot currently be assessed. CONCLUSION(S): Reports based on ASRM/SART Registry data can be enhanced by refined casemix adjustment, assessing outcome rates per patient, as well as per component of ART procedure, and by providing an indication of the statistical significance of observed differences in event rates. In addition, a critical appraisal of available evidence related to particular aspects of infertility management would help clarify the areas in which there is an evidentiary basis for formulation of practice guidelines, as well as topics requiring additional clinical research.

Cryopreservation↗

Comparison of sperm binding potential of uninseminated, inseminated-unfertilized, and fertilized-noncleaved human oocytes under hemizona assay conditions.

In this study, human oocytes obtained after ovarian hyperstimulation for in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT) were utilized to evaluate sperm/zona pellucida binding potential. Three groups of oocytes were evaluated: 1) uninseminated; 2) inseminated-unfertilized; and 3) fertilized-uncleaved. All oocytes had undergone germinal vesicle breakdown at the time of retrieval and were salt-stored (pH 7.2) for not more than 30 days. Sperm binding was recorded under hemizona assay (HZA) conditions using spermatozoa from eight fertile men (HZA control) and from 1) four teratozoospermic (HZA test) and 2) four normozoospermic (HZA test) infertile men. First, the mean numbers (+/- SD) of sperm tightly bound for fertile controls and teratospermic men to hemizonae from uninseminated oocytes were 69.7 +/- 16 and 14.5 +/- 7, respectively (P = 0.02). Likewise, hemizonae from uninseminated oocytes bound 102.0 +/- 19 and 114.0 +/- 28, respectively, for fertile controls and normospermic men (P = 0.5). Second, hemizonae obtained from inseminated-unfertilized IVF oocytes bound 44.2 +/- 12 and 19.7 +/- 6 for fertile controls and teratospermic men, respectively (P = 0.02). This category of oocytes bound 100.5 +/- 7 and 108.5 +/- 11 sperm, respectively, for fertile controls and normospermic semen (P = 0.3). Third, HZA results of fertilized but uncleaved oocytes showed a mean number of tightly bound sperm of 6.0 +/- 4 compared with 65.0 +/- 1 in control, uninseminated oocytes using fertile sperm. These results demonstrate that uninseminated and inseminated-unfertilized human oocytes, salt-stored under controlled pH conditions, give reliable information regarding sperm binding potential under HZA conditions.

Female↗

[Current program of in-vitro fertilization at the Erasmus Hospital: initial results and original ethical aspects].

The clinical results including all in vitro fertilization (IVF) cycles with oocyte pick-up in 1990 are presented. Different types of treatment including classical IVF and embryo transfer, laparoscopic replacement of zygotes in the fallopian tube (ZIFT), IVF with donor sperm (IVF-D), cross fertilization test, embryo freezing, oocyte donation and IVF with epididymal sperm were performed. The total pregnancy rate obtained reaches 38% per oocyte pick-up, 30% of clinical pregnancies (including 4 pregnancies obtained with frozen and thawed embryos). The anticipated "Take Home Baby Rate" will be around 25% per oocyte pick-up, 26 of these 40 pregnancies being today over 20 weeks of gestation. Particular ethical aspects of the program are presented: a study on couple's attitudes regarding embryo freezing as well as the final destination of possibly remaining supernumerary embryos will stress the importance of a precise clear decision on that matter before entering IVF treatment. Indeed the couple's idea on embryo destiny were very precise but also very different. The oocyte donation program has the originality of preserving the donor's anonymity by exchanging the donors recruited by the patients. It will be stressed that this kind of approach combines higher pregnancy chances for the patients, respect of ethical principles linked to gamete donation and gives satisfaction to the patients. The global normalized pregnancy cumulative curve shows that 60% of the couples entering IVF treatment will obtain a child within the first three pick-up cycles.

Embryo Transfer↗