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Cocultures: a new lead in embryo quality improvement for assisted reproduction.

One of the contributory causes to poor PRs in assisted reproduction has been the decreased viability of transferred embryos and the transfer of four-cell embryos into an environment that naturally would be receptive only to 5-day-old blastocysts. In this paper, we have reviewed our own work and that of others on the role of tubal ampullary cells (cocultures) to mimic the in vivo environment to bring about improved embryo quality and an increased number of blastocysts for replacement in IVF patients. The establishment, maintenance, and behavior of human tubal cell lines is first presented, followed by their use as cocultures for fertilization and cleavage of embryos. The mode of action, specificity, and cryopreservation of ampullary cells are also discussed. The currently available results of pregnancies after cocultures are presented together with future aspects of research that are necessary to refine the coculture system. The ultimate aim is to mimic in vivo conditions in vitro, so that at least the PRs of assisted conception can be parallel to normal fecundity in the human. Therefore, a very attractive future includes the freezing of blastocysts generated from coculture, thawing, and replacing them in natural cycles.

Animals↗

Failure of intrauterine insemination in a refractory infertility population.

Intrauterine insemination by itself for multiple and/or severe infertility factors had no benefit over cervical cap with whole ejaculate or coitus in this study. The PRs for IUI and cervical cap with whole ejaculate or coitus were similar and low, suggesting that IUI by itself has limited, if any, utility in enhancing PRs in this type of infertility population. Couples attempting IUI should be advised about the low probability of achieving pregnancy. Ovulation stimulation and/or heterologous donor insemination, IVF, or gamete intrafallopian transfer may be beneficial therapeutic options.

Adult↗

The response of patients with polycystic ovarian disease to human menopausal gonadotropin therapy after ovarian electrocautery or a luteinizing hormone-releasing hormone agonist.

OBJECTIVE: To compare the effect of ovarian electrocautery versus an intranasal (IN) luteinizing hormone-releasing hormone agonist (LH-RH-a) in the response of patients with polycystic ovarian disease (PCOD) to human menopausal gonadotropin (hMG) therapy. DESIGN: A prospective study with serial randomization of patients in two groups for treatment with ovarian electrocautery + hMG or LH-RH-a + hMG. SETTING: A teaching hospital reproductive endocrinology clinic. PATIENTS: Thirty-three women with PCOD who failed to conceive after six treatment cycles with hMG. MAIN OUTCOME MEASURES: Midcycle and luteal phase endocrinology, ovulation, pregnancy rates (PRs), and miscarriage rates. RESULTS: There was no difference in the ovulation or PRs between the two groups. However, the number of cycles with multiple dominant follicles, the luteal phase serum testosterone, and the miscarriage rate were lower in the group pretreated with ovarian electrocautery. CONCLUSIONS: Pretreatment of patients with PCOD with ovarian electrocautery may be a better alternative to IN LH-RH-a therapy for induction of ovulation with hMG.

Administration, Intranasal↗

The appearance of one-pronuclear human oocytes is associated with a better ovulation-induction response and successful pregnancy outcome.

OBJECTIVE: To study the relationship between the presence of one-pronuclear oocytes in in vitro fertilization (IVF) patients and ovulation-induction response, oocyte and embryo development, and clinical outcome. DESIGN: Retrospective analysis of 535 consecutive IVF retrievals. Retrievals in which one or more oocytes exhibited one pronucleus were compared with retrievals in which no one-pronuclear oocytes (control) were observed. The following one-pronuclear versus control subgroups were also examined: leuprolide acetate/human menopausal gonadotropin (LA/hMG) ovulation inductions, high estradiol (E2) response cases, and retrievals in which a large number of oocytes (greater than or equal to 15) were recovered. SETTING: Brigham and Women's Hospital, a tertiary care, university-affiliated hospital. PATIENTS: Three hundred forty-six IVF patients were treated between January 1989 and May 1991. MAIN OUTCOME MEASURES: Parameters examined included E2 concentration and number of follicles with maximum diameter greater than or equal to 12 mm on day of human chorionic gonadotropin administration; number of total and mature oocytes retrieved; total fertilization rates; number of embryos; and percent per retrieval of embryo transfers (ETs), clinical pregnancies, and ongoing-livebirths. RESULTS: The one-pronuclear patients had higher E2 levels and larger number of follicles, yielded significantly more total and mature oocytes, had a higher overall fertilization rate, produced more embryos, and had higher ET, clinical pregnancy and ongoing-livebirth rates per retrieval than did the control patients. Analysis of the subgroup populations revealed no significant differences in the majority of the main outcome measures studied; however, the one-pronuclear patients yielded significantly more total and mature oocytes per retrieval. CONCLUSIONS: Although there was an increase in the clinical and ongoing-livebirth pregnancy rates (PRs) in one-pronuclear patients, this was probably associated with an improved ovulation-induction response in the one-pronuclear patients. They achieved significantly higher E2 levels, recruited a larger number of follicles, and yielded more oocytes and embryos per retrieval than the control patients. When only the LA/hMG, E2 greater than or equal to 1,500 pg/mL, or the greater than or equal to 15 oocytes/case retrievals were analyzed, the PRs were no longer different; however, the one-pronuclear patients still yielded significantly more total and mature oocytes per retrieval than the controls. Therefore, the appearance of one-pronuclear oocytes is probably associated with the maturation stage of the oocytes obtained and is indicative of an ovulation induction in which a large number of preovulatory, metaphase II oocytes have been recruited.

Cell Nucleus↗

Exogenous control of the cycle simplifies thawed embryo transfer and results in a pregnancy rate similar to that for natural cycles.

To determine if a satisfactory PR could be achieved in thawed ET using a simplified protocol, standard monitoring in the natural cycle was compared with a controlled preparation of the endometrium with exogenous E2 and P without hormonal or ultrasonographic monitoring. All patients had normal ovarian function, and no GnRH-a were administered. Pregnancy rates, ongoing PRs, and embryo implantation rates were similar in the two groups. The use of exogenous E2 and P without GnRH-a suppression to control the cycle for thaw ET is safe, convenient, and results in PRs at least as good as in the natural cycle. It may be of particular value in patients with an irregular cycle or in those for whom prolonged monitoring is undesirable.

Cryopreservation↗

Comparison of results obtained with human serum and a protein solution as a supplement for in vitro fertilization culture medium.

A prospective controlled study was performed to compare the PRs obtained after use of a uniform IVF culture medium containing a pasteurized serum protein solution or patient serum. The ongoing PRs per ET in the serum and the protein solution group were 32% and 28%, respectively (not significant). Culture of supernumerary embryos showed blastocyst formation and even hatching with both supplements. The PR will not drop when this protein solution is used as a protein supplement in IVF culture medium instead of patient serum.

Blood↗

Freezing cocultured human blastocysts.

OBJECTIVE: To determine the possibility of obtaining good pregnancy rates (PRs) after freezing and thawing cocultured blastocysts. DESIGN: Human blastocysts were frozen first according to a protocol available from literature. Two other protocols including the addition of glycerol were designed to improve the results. SETTING: All the patients were under clinical management at the Institut Rhonalpin pour l'Etude de la Reproduction Humaine in Lyons, France. PATIENTS: Patients involved in the in vitro fertilization program have had their supernumerary embryos frozen according to the three protocols presented here. MAIN OUTCOME MEASURES: Embryo recovery after freezing and thawing. Clinical and ongoing pregnancies after embryo transfer (ET). RESULTS: A protocol including sucrose addition and reduction of steps in the preparation of the blastocysts for freezing gave us a 21% PR per transfer (15 ongoing) of 101 transfers (106 thawings). CONCLUSIONS: Freezing of cocultured human blastocysts allow good PRs. This can represent an alternative for repeated failures of ETs at early stages.

Blastocyst↗

The relative success of gonadotropin-releasing hormone analogue, clomiphene citrate, and gonadotropin in 1,099 cycles of in vitro fertilization.

OBJECTIVES: To evaluate the effectiveness of and analyze the factors influencing the outcome of three ovarian stimulation protocols used during in vitro fertilization (IVF) in a large population. DESIGN: Retrospective file review. SETTING: In vitro fertilization program in one center during the years 1985 to 1990. PATIENTS AND PROTOCOLS: Three hundred forty-one patients received clomiphene citrate (CC) and human menopausal gonadotropin (hMG), 365 received hMG alone, and 393 received gonadotropin-releasing hormone analogue (GnRH-a) for pituitary suppression followed by hMG stimulation. MAIN OUTCOME MEASURE: Rates of cancellation, total pregnancies, and ongoing pregnancies, with breakdown by age of patients. RESULTS: The cancellation rate because of early luteinization following GnRH-a/hMG was significantly reduced compared with the other two protocols: 3.6% versus 9.4% and 13.7% for CC/hMG and hMG, respectively. However, in women over 40 years of age, GnRH-a/hMG resulted in the highest rate of poor ovarian response. Significantly more oocytes were retrieved, fertilized, and cleaved after the use of GnRH-a/hMG compared with the other two protocols. Despite this, clinical pregnancy rate (PR) was the highest with CC/hMG compared with GnRH-a/hMG and hMG:31.4% versus 16.9% and 15.7%, respectively. Ongoing PRs were 20.5%, 9.7%, and 11.6%, respectively. CONCLUSIONS: Although the use of GnRH-a for pituitary suppression before ovarian stimulation for IVF reduced the cancellation rate and increased the number of retrieved oocytes, it was not found to result in higher PRs than those achieved by stimulation with CC/hMG. This suggests that treatment by GnRH-a/hMG should be reserved mainly for the prevention of early luteinization.

Clomiphene↗

The effect of endometrial thickness and echo pattern on in vitro fertilization outcome in donor oocyte-embryo transfer cycle.

There have been some conflicting data concerning the importance of endometrial thickness and echo patterns before transfer in different IVF-ET situations under different COH regimens. We previously found in women undergoing IVF-ET after luteal phase LA-hMG a significantly higher PR in those patients attaining at least a 10-mm endometrial thickness and a lower rate in those women with an entirely homogeneous hyperechogenic endometrium (pattern C). The present study evaluated the relationship of endometrial thickness and echo pattern to PRs in donor oocyte recipient immediately before transfer. There were 16 pregnancies in 58 cycles (27.5%). Conclusions similar to the previous COH study were reached concerning the > or = 10-mm thickness levels correlating with improved PRs (9% versus 38.7%, P < 0.01). In contrast, no correlation with echo pattern was found.

Adult↗

The role of aerobic and anaerobic semen cultures in asymptomatic couples undergoing in vitro fertilization: effects on fertilization and pregnancy rates.

OBJECTIVE: To determine if routine semen culture is useful in asymptomatic couples undergoing in vitro fertilization and embryo transfer (IVF-ET). DESIGN: Prospective data collection. SETTING: All cultures and IVF cycles were performed at the University of North Carolina in Chapel Hill, North Carolina. PARTICIPANTS: All asymptomatic couples undergoing IVF-ET from January 1989 through January 1990. INTERVENTIONS: Aerobic and anaerobic cultures were performed on semen samples obtained before IVF. MAIN OUTCOME MEASURES: Quantitative semen cultures were evaluated for both aerobic and anaerobic bacterial isolates. Fertilization and pregnancy rates (PRs) were compared in patients with positive and negative semen cultures. RESULTS: Eighty percent of cultures contained at least one bacterial isolate. Three of the four most commonly isolated bacteria were normal skin flora. Positive culture results had no effect on either fertilization or PRs. CONCLUSIONS: Bacterial contamination is common with semen collection, yet routine semen cultures are not beneficial in asymptomatic couples undergoing IVF-ET.

Adult↗

Comparison of short 7-day and prolonged treatment with gonadotropin-releasing hormone agonist desensitization for controlled ovarian hyperstimulation.

OBJECTIVE: To compare two treatment regimens associating a gonadotropin-releasing hormone agonist (GnRH-a) and human menopausal gonadotropin (hMG) for controlled ovarian hyperstimulation (COH). DESIGN: A prospective randomized trial. SETTING: The outpatient fertility clinic of a university tertiary care center, the Hôpital A. Béclère, Clamart, France. PARTICIPANTS: One hundred eighty-two in vitro fertilization (IVF) candidates undergoing new or repeat IVF cycles at Hôpital A. Béclère over a 4-month period. TREATMENT: Group 1 (7-day protocol): A short-acting preparation of GnRH-a (Tripteriline 0.1) was administered daily for 7 days, starting on cycle day 2. Ovarian stimulation with hMG was started on cycle day 4. Group 2 (long protocol): A timed release preparation of GnRH-a (Tripteriline 3.75 mg) was administered on cycle day 2. Ovarian stimulation with hMG was started after documented ovarian suppression. MAIN OUTCOME MEASURES: Response to COH, pregnancy rate (PR), tolerance. RESULTS: In the 7-day protocol, the amount of hMG required was markedly lower at 24 +/- 7 than in the long protocol group requiring 42.5 +/- 9.75 vials (75 IU) (mean +/- SD). No elevation of plasma LH occurred in either group. The number of oocytes retrieved was 7.3 +/- 1 and 10.7 +/- 1.2 (mean +/- SD) in the 7-day and long protocols, respectively. Yet, the number of embryos obtained and the PRs were similar in the two treatment groups. CONCLUSIONS: We observed that in COH, GnRH-a treatment could be interrupted safely several days before human chorionic gonadotropin administration without risking a premature increase of plasma luteinizing hormone. Moreover, the number of embryos available for fresh transfer and the ongoing PRs were similar in the new 7-day and in the classic long GnRH-a/hMG protocols, despite the smaller number of oocytes suggesting a greater efficiency of the 7-day protocol. The peak estradiol level and the hMG requirement were also lower in the 7-day GnRH-a/hMG protocol.

Adult↗

Transvaginal versus laparoscopic gamete intrafallopian transfer: a case-controlled retrospective comparison.

OBJECTIVE: To compare pregnancy rates (PRs) obtained by transvaginal ultrasound (US)-guided GIFT with PRs from conventional laparoscopy-based GIFT. DESIGN: Retrospective case-controlled study. SETTING: Large, private assisted conception service. PATIENTS: Twenty consecutive cycles of transvaginal GIFT, each matched with three cycles of laparoscopic GIFT on the basis of patient age, number of previous unsuccessful attempts at GIFT, number of eggs transferred, and total number of eggs obtained. MAIN OUTCOME MEASURE: Probability of clinical pregnancy. RESULTS: The PR from transvaginal GIFT was 20.0% (95% confidence limits 13% to 27%), compared with a PR from laparoscopic GIFT of 35.0% (23% to 50%). CONCLUSION: Transvaginal GIFT with present techniques seems to be less effective than laparoscopic GIFT, but it can be considered as a practical alternative to ovarian stimulation and intrauterine insemination and to peritoneal ovum-sperm transfer.

Adult↗

Fertility after ectopic pregnancy.

OBJECTIVES: To compare pregnancy rates (PRs) after radical or conservative surgical treatment for tubal pregnancy over a 12.5-year interval (minimum of 3 years) and to assess the relative contribution of various risk factors to future fertility performance. DESIGN: A retrospective cohort study examining the influence of various risk factors on PRs with stepwise discriminant analysis using a jackknife-type validation program. PATIENTS: Eighty-eight Olmsted County, Minnesota, women who presented over a 10-year interval with their first ectopic pregnancy (EP), undergoing either radical or conservative surgery at laparotomy, who actively attempted conception after surgery and who were available for follow-up at Mayo Clinic. MAIN OUTCOME MEASURES: The primary end point for analysis was the occurrence of a livebirth or EP at 3 years of follow-up after the index EP. RESULTS: No difference in PR was identified in patients treated with radical or conservative surgery. Patients who underwent conservative surgery had a higher risk of subsequent EP. Patients with a prior history of infertility had significantly reduced fertility potential. CONCLUSIONS: Prior history of infertility was the most significant determinant for fertility potential after surgical treatment for EP. Patients with prior infertility had markedly impaired fertility after surgical treatment of EP, and outcome was not influenced by choice of surgical procedures. Patients without prior infertility had comparable encouraging fertility potential with both conservative and radical therapy. These findings have relevance for future management of infertility in affected patients.

Discriminant Analysis↗

Endometrial receptivity: the age-related decline in pregnancy rates and the effect of ovarian function.

OBJECTIVE: To assess the effect of age and ovarian function on endometrial receptivity. DESIGN: Retrospective comparison between standard IVF and ovum donation in younger and older patients (< 40 and > or = 40 years of age, respectively). PATIENTS: In standard IVF, there were 325 transfer cycles in older patients and 1,103 transfer cycles in younger ones. In ovum donation, there were 236 transfer cycles in older patients and 222 cycles in younger women. Ovum recipients were then redivided into two groups, according to ovarian function: ovarian failure group (219 cycles) and eugonadal group (239 cycles) in patients with retained ovarian function as manifested by regular menstrual cycles and normal gonadotropins. RESULTS: In standard IVF, clinical pregnancy rates (PRs) were significantly lower in older patients (12.9% versus 23.8%, respectively). In ovum donation, clinical PRs were also significantly lower in older patients (21.2% versus 29.3%, respectively). A significantly higher clinical PR (31.1%) was noted in patients with ovarian failure, compared with both eugonadal patients undergoing ovum donation (19.7%) and standard IVF patients (21.3%). CONCLUSIONS: The decrease in endometrial receptivity with age is responsible for the higher rate of implantation failure in older women. Patients with nonfunctioning ovaries do better than eugonadal patients in ovum donation programs.

Adult↗

Pregnancy rates in donors versus recipients according to the serum progesterone level at the time of human chorionic gonadotropin in a shared oocyte program.

Previous data suggested a subtle increase in serum P at the time of hCG injection without LH surge reduces the PR of women having oocyte retrievals for IVF; this study compared PRs of recipients in a shared oocyte program according to the donors' pre-hCG P level. There was no difference in viable PRs between recipients and donors when P < or = 1 ng/mL. The PR was similar for recipients when donors' P was > 1 ng/mL (12.7%). Donors with P > 1 ng/mL had the lowest PR: 7.2%. The data suggest that the adverse effect of higher serum P without LH surge may be on the endometrium rather than the oocyte.

Chorionic Gonadotropin↗

Prospective study of a modified gonadotropin-releasing hormone agonist long protocol in an in vitro fertilization program.

OBJECTIVE: To determine if pituitary suppression is still maintained if GnRH agonist (GnRH-a) is discontinued as gonadotropin stimulation is begun in a long protocol. DESIGN: Prospective, randomized study. SETTING: An outpatient IVF-GIFT program. PATIENTS: One hundred seventy-three patients entering an IVF-GIFT program. INTERVENTIONS: Gonadotropin-releasing hormone agonist in long protocol was either discontinued or continued as gonadotropin stimulation was begun. MAIN OUTCOME MEASURES: Luteinizing hormone (LH), E2, and P levels, egg numbers, fertilization rate, number of embryos transferred, day of gonadotropin stimulation, and pregnancy rates (PRs). RESULTS: Pituitary suppression was maintained although GnRH-a was discontinued as gonadotropin stimulation was begun. No spontaneous LH surge was seen, and PRs were increased in the IVF patients. CONCLUSION: This study indicates that the advantages gained from use of GnRH-a in the long protocol are not compromised by its early discontinuation.

Adult↗

The role of estrogen support during the luteal phase of in vitro fertilization-embryo transplant cycles: a comparative study between progesterone alone and estrogen and progesterone support.

OBJECTIVE: To evaluate the possible role for estrogen supplementation to the P luteal phase support of GnRH agonists (GnRH-a)- and hMG-induced IVF-ET cycles. SETTING: In vitro fertilization unit in a tertiary care university hospital. DESIGN: A prospectively randomized study. PATIENTS: One hundred consecutive patients undergoing ET after IVF were assigned into one of two luteal supplementation regimens. INTERVENTIONS: In all patients enrolled in the study, ovulation was induced using the midluteal regimen for pituitary down regulation with GnRH-a followed by follicular stimulation with hMG. The first group received IM P 50 mg/d, as luteal phase support, starting the day of ET. The second group received the same dosage of P, combined with oral E2 valerate, 2 mg/d. Serum levels of P and E2 were monitored every 4 days for 16 days after ET. MAIN OUTCOME MEASURES: Pregnancy rates (PRs) and live birth rates per ET. RESULTS: No significant difference in E2 or P levels throughout the cycle was observed between groups. Similar PRs per ET and the live birth rates were also observed between group A and B (28% versus 26.5% and 78.6% versus 76.1%, respectively). CONCLUSION: No advantage was found in the addition of E2 valerate to P luteal phase support of GnRH-a- and hMG-induced IVF-ET cycles.

Adult↗

A randomized, double-blind, placebo-controlled study of luteal phase dydrogesterone (Duphaston) in women with minimal to mild endometriosis.

OBJECTIVE: To compare dydrogesterone with placebos in the treatment of minimal to mild endometriosis. DESIGN: Prospective, double-blind, randomized study. SETTING: Three Obstetrics and Gynaecology Departments in the United Kingdom. PATIENTS: Sixty-two premenopausal women with complaints of pain (n = 12) and infertility with or without pain (n = 50) with minimal to mild endometriosis diagnosed at laparoscopy. Thirty-nine women had a laparoscopy after treatment and 56 women were followed up 12 months after treatment. INTERVENTIONS: Two high doses of dydrogesterone (either 40 or 60 mg) or a placebo, which was given for 12 days, beginning 2 days after the LH surge for a treatment period of 6 months. MAIN OUTCOME MEASURES: Change between before and after treatment endometriosis scores, pregnancy rates (PRs), and pain. RESULTS: Treatment with dydrogesterone did not alter the natural history of endometriosis or PRs when compared with placebo. Pain was reduced significantly during treatment with 60 mg dydrogesterone and this improvement still was evident at 12-month follow-up. CONCLUSION: Luteal phase dydrogesterone reduces pain associated with endometriosis.

Adult↗