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PubMed2006

Effect of endometrial cavity fluid on clinical pregnancy rate in tubal embryo transfer (TET).

PURPOSE: In this study, we assess whether the endometrial cavity fluid (ECF) generated physiologically by the genital tract have negative effect on the pregnancy rate during tubal embryo transfer (TET) in patients who do not have hydrosalpinx or bilateral tubal obstruction. METHODS: We retrospectively collected samples from 176 women with 195 cycles receiving TET due to male factor, unexplained infertility or endometriosis from June 1999 to Dec. 2003, and divided them into two groups (group I: patient with fluid accumulation >1 mm in the anterior-posterior diameter in the uterine cavity; group II: patient without fluid accumulation in the uterine cavity). Endometrium thickness was measured as a maximal distance between anterior and posterior myometrium-endometrium interface under the long-axis view. The A-P diameter of ECF was measured via vaginal sonar on the day of ovum pick-up (OPU). RESULTS: From a total of 195 ART cycles involving 176 patients, the accumulation of ECF was detected in 10 cycles (5.1%). Seven of ten cycles (70%) with the accumulation of ECF were proved to be pregnant clinically. However, in the rest 185 cycles (group II), 98 of them (53%) were proved to be pregnant. At the mean time, the implantation rate was 29.03% and 25.71% in the groups I and II, respectively. No significant difference of the clinical pregnancy rate and the implantation rate was found between the two groups. CONCLUSIONS: According to our study, if the ECF was generated physiologically by the genital tract during controlled ovarian hyperstimulation (COH), the clinical pregnancy rate is not worse and no influence of embryo implantation was found.

Adult↗
PubMed1994

Bovine nuclear transfer embryos: oocyte activation prior to blastomere fusion.

Successful bovine nuclear transfer (NT) embryo production requires proper oocyte activation and transfer of a nucleus into this oocyte. However, the temporal relationship between these two events is unclear. The current study examined whether activation of the oocyte prior to fusion would induce nuclear swelling while also affecting development to morula and blastocyst stage and finally development to offspring. Aged oocytes can be activated by a number of techniques including exposure to room temperature. In this study oocyte activation was induced through three different means: reduced temperature culture alone, reduced temperature culture and calcium ionophore, and naturally, through the fertilization process. Electrofusion was carried out after the activation stimulus. When used in the NT procedure, activation of oocytes prior to fusion resulted in NT embryos that underwent nuclear swelling and had a high developmental rate to morula and blastocyst stages. Also, these NT embryos developed to normal offspring when transferred to recipient animals. The addition of a calcium ionophore treatment to the reduced temperature culture was not beneficial and resulted in less nuclear swelling. The use of enucleated fertilized oocytes as recipient cytoplasm for the new nucleus resulted in NT embryos developing to morula and blastocyst stages at the same rate as room temperature activated NT embryos. Therefore, improved embryo development can be obtained from NT embryos if the aged recipient oocyte is activated prior to the time of fusion. Also, offspring were obtained from these pre-activated NT embryos.

Animals↗
PubMed2004

Comparison of two different starting multiple dose gonadotropin-releasing hormone antagonist protocols in a selected group of in vitro fertilization-embryo transfer patients.

OBJECTIVE: To compare the efficacy of two starting protocols of multiple dose GnRH antagonists (GnRH-a). DESIGN: Prospective randomized controlled study. SETTING: In vitro fertilization-embryo transfer program at the Instituto Valenciano de Infertilidad, Valencia, Spain. PATIENT(S): One hundred nine patients undergoing controlled ovarian hyperstimulation (COH) with recombinant gonadotropins and GnRH-a (0.25 mg/d). INTERVENTION(S): Patients started GnRH-a administration on stimulation day 6 (group 1) or when the leading follicle reached a mean diameter of 14 mm (group 2). MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates; serum E(2) and LH levels during ovarian stimulation; days of stimulation and GnRH-a administration. RESULT(S): Days needed for ovarian stimulation were similar in both groups but there was a significant difference when comparing days of GnRH-a administration. Serum E(2) and LH followed similar curves in both groups. Implantation and pregnancy rates were 23.7% and 44.4 % in group 1 and 28.6% and 50.9 % in group 2 (P=not significant [NS]). CONCLUSION(S): The efficacy of the two starting protocols of the multiple dose GnRH-a evaluated in this study is similar; however, this remark can only be drawn for a selected group of patients.

Adult↗
PubMed2001

Does prayer influence the success of in vitro fertilization-embryo transfer? Report of a masked, randomized trial.

OBJECTIVE: To assess the potential effect of intercessory prayer (IP) on pregnancy rates in women being treated with in vitro fertilization-embryo transfer (IVF-ET). STUDY DESIGN: Prospective, double-blind, randomized clinical trial in which patients and providers were not informed about the intervention. Statisticians and investigators were masked until all the data had been collected and clinical outcomes were known. The setting was an IVF-ET program at Cha Hospital, Seoul, Korea. IP was carried out by prayer groups in the United States, Canada and Australia. The investigators were at a tertiary medical center in the United States. The patients were 219 women aged 26-46 years who were consecutively treated with IVF-ET over a four-month period. Randomization was performed after stratification of variables in two groups: distant IP vs. no IP. The clinical pregnancy rates in the two groups were the main outcome measure. RESULTS: After clinical pregnancies were known, the data were unmasked to assess the effects of IP after assessment of multiple comparisons in a log-linear model. The IP group had a higher pregnancy rate as compared to the no-IP rate (50% vs. 26%, P = .0013). The IP group showed a higher implantation rate (16.3% vs. 8%, P = .0005). Observed effects were independent of clinical or laboratory providers and clinical variables. CONCLUSION: A statistically significant difference was observed for the effect of IP on the outcome of IVF-ET, though the data should be interpreted as preliminary.

Adult↗
PubMed2004

Preimplantational embryo development and incidence of blastomere apoptosis in bovine somatic cell nuclear transfer embryos reconstructed with long-term cultured donor cells.

This study was performed to investigate whether types and/or age of donor cells affect preimplantational embryo development and the incidence of apoptosis in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine fetal or adult ear fibroblasts were isolated, cultured in vitro and categorized into fresh or long-term cultured cells in terms of population doublings (PD): in fetal fibroblasts, <16 being considered fresh and >50 being long-term cultured; in adult ear fibroblasts, <16 being considered fresh and >30 being long-term cultured. Bovine oocytes from slaughterhouse ovaries were matured in TCM-199, enucleated and reconstructed by SCNT. The reconstructed oocytes were fused, chemically activated, and cultured in modified synthetic oviduct fluid (mSOF) at 39 degrees C in a humidified atmosphere of 5% CO(2) air for 7 days. The early development of SCNT embryos was monitored under a microscope and the quality of blastocysts was assessed by differential counting of inner cell mass (ICM) and trophectoderm (TE) cells and by apoptosis detection in blastomeres using a terminal deoxynucleotidyl transferase-mediated d-UTP nick end-labeling (TUNEL) assay. As results, types and/or age of donor cells did not affect the rate of blastocyst formation and the number of ICM and TE cells. However, a significant increase in apoptotic blastomeres was observed in SCNT embryos reconstructed with long-term cultured fetal or adult ear fibroblasts compared to those in SCNT embryos derived from fresh fetal or adult ear fibroblasts. In conclusion, these results indicated that the long-term culture of donor cells caused increased the incidence of apoptosis in bovine SCNT embryos but did not affect the developmental competence and the cell number of blastocysts.

Animals↗
PubMed1999

Assessment of endometrial volume by three-dimensional ultrasound prior to embryo transfer: clues to endometrial receptivity.

The aim of the present prospective study was to obtain quantitative data on endometrial volume by three-dimensional (3D) ultrasound at the time of embryo transfer in an in-vitro fertilization programme and to assess its value in predicting endometrial receptivity. The cycles (n = 72) were classified according to endometrial volume: group A <2 ml, group B 2-4 ml, and group C >4 ml. Comparisons of the groups showed that pregnancy and implantation rates were significantly lower (P < 0.05) in the group of patients with an endometrial volume <2 ml. Furthermore, no pregnancy was achieved with an endometrial volume <1 ml. It is concluded that endometrial volume by 3D transvaginal ultrasound may become a new objective parameter by which to predict endometrial receptivity.

Adult↗
PubMed1984

Delayed appearance of plasmatic chorionic gonadotropin in pregnancies after in vitro fertilization and embryo transfer.

The detection of pregnancy through the rise of human chorionic gonadotropin hormone secretion, on maternal plasma level, has been studied in normally developed pregnancies following in vitro fertilization and embryo transfer (IVF-ET), and compared with two other groups of pregnancies, the first group being pregnancies following artificial insemination with donor semen (AID) in spontaneous cycles ("AID group") and the second group being pregnancies following in vivo fertilization in a stimulated cycle ("stimulated group"). The day of human chorionic gonadotropin detection level significant for pregnancy (Dd) has been first defined and then determined for each pregnancy. Thereafter, mean levels for Dd (Dd) have been compared for each pregnancy group. It has been found that in pregnancies following IVF-ET, Dd is 12.05 +/- 0.8 days after ovulatory stimulus, which is delayed in comparison with spontaneous cycle pregnancies (Dd = 9.5 +/- 1.0) and with stimulated cycle pregnancies (Dd = 8.0 +/- 1.5). The hypothesis to explain this observation is then discussed.

Adult↗
PubMed1999

Endovaginal color Doppler sonographic evaluation of ectopic pregnancy in women after in vitro fertilization and embryo transfer.

The aim of this study was to evaluate the value of endovaginal color Doppler ultrasonography in the early diagnosis of ectopic pregnancy in women after in vitro fertilization and embryo transfer, and to correlate the sonographic findings with ss-hCG serum levels. Thirty-five patients had proven ectopic pregnancies and 4 other patients had heterotopic pregnancies. The diagnosis was disclosed correctly in all cases by endovaginal color Doppler US by identifying an adnexal mass with placental flow and a nongravid uterus called a "cold uterus". An intrauterine sac with "double ring sign" was found in all normal intrauterine pregnancies when the hCG levels exceeded 1000 IU/l but in none of the patients with ectopic pregnancy (EP). These findings suggest the efficacy of the discriminatory hCG serum level of 1000 IU/l in the investigation of EP. In conclusion, this study describes the diagnostic importance of transvaginal color Doppler US in correlation with hCG serum levels in the early detection of EP avoiding life-threatening complications and improving patient outcome.

Adult↗
PubMed1992

Direct enzyme immunoassay of estradiol in serum of women enrolled in an in vitro fertilization and embryo transfer program.

We present a fast and simple direct enzyme immunoassay for the sexual steroid hormone estradiol-17 beta in serum of women enrolled in an in vitro fertilization and embryo transfer program. We added 25 microL of standards or samples and 125 microL of a mixture of monoclonal antibody and displacing agents to the wells of a microtiter plate previously coated with a second antibody. After 30 min 50 microL estradiol-horseradish peroxidase conjugate is added. Total incubation time is 75 min. The immunoassay is stopped by washing the microtiter plate. The amount of bound conjugate is then measured at 450 nm with hydrogen peroxide as a substrate and 3,3',5,5'-tetramethylbenzidine as chromogen. The detection limit is 0.24 nmol/L. The assay is thus limited in its application. Analytical recovery is 90.8% (range 80.4-102.9%); within- and between-assay CVs range between 1.3% and 15.4%. Total assay time, including preparation of reagents and data reduction, is only 2.5 h. Results of this enzyme immunoassay compare well with those obtained with four different commercial radioimmunoassays (r = 0.88-0.93).

Embryo Transfer↗
PubMed1993

Eliminating the risk of life-endangering complications following overstimulation with menotropin fertility agents: a report on women undergoing in vitro fertilization and embryo transfer.

OBJECTIVE: To evaluate a new method for preventing the life-endangering complications associated with inadvertent menotropin-induced severe ovarian hyperstimulation in patients undergoing in vitro fertilization and embryo transfer (IVF-ET). METHODS: Seventeen women each underwent a single cycle of controlled ovarian hyperstimulation with menotropins in preparation for IVF-ET. The indications for IVF-ET were tubal occlusion in nine, endometriosis in six, and unexplained infertility in two. The peak plasma estradiol (E2) concentration before hCG administration was greater than 6000 pg/mL and more than 30 ovarian follicles were detected by transvaginal ultrasound. Thus, life-endangering complications associated with severe ovarian hyperstimulation syndrome were highly likely to occur following hCG administration. Rather than cancel the cycle of treatment, menotropin therapy was discontinued and hCG administration was deferred for a number of days until the plasma E2 concentration fell below 3000 pg/mL ("prolonged coasting"), whereupon hCG was administered and egg retrievals and ETs were duly performed. RESULTS: None of the women developed severe ovarian hyperstimulation syndrome. There were six viable pregnancies (35.2%), which proceeded normally. CONCLUSION: This study indicates that "prolonged coasting" prevents severe ovarian hyperstimulation syndrome in severely overstimulated women undergoing IVF-ET, without necessitating cycle cancellation.

Adult↗
PubMed2005

[Study on the correlation of transforming growth factor beta1 and its receptors with spontaneous abortion after in vitro fertilization and embryo transfer].

OBJECTIVE: To understand the expressions of transforming growth factor beta1 (TGF-beta1) and its receptors (TbetaR) in villi and decidua during early pregnancy and their effects on early embryo development, and explore the mechanism of spontaneous abortion. METHODS: By immunohistochemical technique, expressions of TGF-beta1 and its receptors were determined in villi and decidua from 10 cases of spontaneous abortion women after in vitro fertilization and embryo transfer (IVF-ET group), 20 cases of spontaneous abortion women (spontaneous abortion group), and 20 cases of normal early pregnancy women (control group). RESULTS: TGF-beta1 and its receptors had high expressions in decidua cells, villus glands and interstitial cytoplasm in all three groups. The average light density of the expressions of TGF-beta1 in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.167 and 0.199, 0.198 and 0.201, 0.277 and 0.274, respectively. The intensity of TbetaR-I in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.144 and 0.150, 0.202 and 0.201, 0.238 and 0.281, respectively. And the expressions of TbetaR-II in the villi and decidua of IVF-ET, spontaneous abortion and control groups were 0.199 and 0.145, 0.153 and 0.156, 0.300 and 0.301, respectively. The differences between the control group and both abortion groups were all significant (P < 0.01). CONCLUSIONS: The expressions of TGF-beta1 and its receptors in the early pregnancy tissues of aborted women declined. It demonstrates that the abnormal expressions of TGF-beta1 and its receptors play an important role in spontaneous abortion, they are one of the causes of spontaneous abortion.

Abortion, Spontaneous↗
PubMed1991

Early pituitary desensitization and ovarian suppression with leuprolide acetate is associated with in vitro fertilization-embryo transfer success.

OBJECTIVE: To determine if the timing of the onset of pituitary desensitization and ovarian suppression using follicular phase leuprolide acetate (LA) is associated with in vitro fertilization-embryo transfer (IVF-ET) success for pregnancy. DESIGN: Retrospective series of IVF patients undergoing pituitary desensitization and ovarian suppression before beginning controlled ovarian hyperstimulation for IVF-ET. SETTING: Tertiary infertility practice. PATIENTS: Seventy-eight women for 80 cycles began LA on day 1 of their menstrual cycle. After 11 days of LA, 47 (59%) cycles in group I had suppressed serum estradiol (E2) levels less than 40 pg/mL, in contrast to 33 (41%) cycles in group II not adequately suppressed, thereby requiring additional days to achieve suppression. INTERVENTIONS: Controlled ovarian hyperstimulation was started when patients were satisfactorily suppressed, i.e., E2 less than 40 pg/mL. MAIN OUTCOME MEASURE(S): Mean E2 response, ampules of human menopausal gonadotropin, cancellation rates, number of oocytes retrieved, fertilization rates, and pregnancy rates (PRs) per cycle were examined between groups I and II. RESULTS: Group I demonstrated a greater mean E2 response on the day of human chorionic gonadotropin 1,735 pg/mL versus 1,470 pg/mL (P = 0.008), a greater fertilization rate 64% versus 55% (P = 0.02), and a higher PR per cycle 34% versus 12% (P = 0.036) compared with group II. CONCLUSIONS: Women who achieved desensitization-suppression within 11 days of initiating LA demonstrated a more favorable outcome for IVF-ET than those who did not.

Chorionic Gonadotropin↗
PubMed1997

[Basic ovarian status and follicular response to superovulation stimulation in an in vitro fertilization and embryo transfer program].

OBJECTIVE: To study the relationship between patient age, cycle day 3 basal ovarian status, serum estradiol (E2) level and the ovarian response in an in vitro fertilization and embryo transfer (IVF-ET) program. METHOD: 102 cases and 102 cycles of IVF-ET patients with regular menstrual period and normal cycle day 3 basal follicular-stimulating hormone (FSH < 20 IU/L) level were studied. The same superovulation regimen was employed. The ovarian response was classified as low when follicle number (diameter > 10 mm) was fewer than 3 on the day of hCG injection, moderate when the number was 3-14, and high when follicle number exceeded 14. RESULTS: (1) Patients older than 35 years tended to be low responders; women younger than 30 years usually responded well with production of more than 15 follicles. (2) The number of cycle day 3 follicles was positively correlated with the number after stimulation with gonadotropin. When total basal follicle number in both ovary exceeded 20, ovarian an hyperstimulation syndrome should be watched out. (3) On cycle day 3 the diameter of the largest follicle was negatively correlated with the ovarian response. In low responders the diameter of the largest follicle was usually larger than 4 mm. (4) Despite of the different size and number of the cycle day 3 follicles the serum E2 level was quite similar. CONCLUSION: Age and basal ovarian status including the number and size of the antral follicles are valuable factors to be considered in the prediction of ovarian response to the same gonadotrophic stimulation protocol.

Adult↗
PubMed1990

Correlation between follicular fluid content and the results of in vitro fertilization and embryo transfer. II. Inhibin and aromatase inhibitor activity.

The inhibin content and aromatase inhibitor activity (AIA) of 72 follicular fluids (FF) obtained from 42 women undergoing in vitro fertilization (IVF) and embryo transfer (ET) were studied as a function of IVF ET outcome. Inhibin levels were determined by bioassay (BA) and RIA; AIA was measured by BA. The inhibin content of follicles characterized as immature by their estradiol (E2) levels and E2/progesterone (P) ratios was significantly lower (P less than 0.05) than that of mature follicles (i.e. leading to pregnancy). The mean AIA for mature follicles were significantly lower than AIA in groups where pregnancy was not obtained. AIA for follicles from which a pregnancy was obtained for each ET was also significantly lower than that in FF characterized as immature of hypermature. The highest E2/AIA and inhibin BA/AIA ratios were associated with the highest incidence of successful IVF ET outcome. No correlation was found between AIA and inhibin, on the one hand, and E2, delta 4-androstenedione, E2/P, and PRL, on the other. However, a positive correlation was found between inhibin (RIA and BA) and P, reflecting the production of inhibin by granulosa cells during luteinization. These studies allowed us to conclude that FF inhibin levels do not differ according to IVF ET outcome, but are an index of follicular maturation. AIA not only constitutes an index of follicular maturation and granulosa cell luteinization, but is of predictive value for IVF ET outcome as E2/AIA and inhibin BA/AIA ratios.

Adult↗
PubMed2006

Effects of maturational age of recipient oocytes and activation conditions on the development of porcine fetal fibroblast nuclear transfer embryos.

This study was conducted to evaluate the nuclear remodeling patterns and the developmental potential of porcine fetal fibroblast nuclear transfer embryos (NTs) following the maturational age of recipient oocytes and activation conditions. Donor cells were transferred into the enucleated oocytes that were matured for 36 or 44h. Electrofused embryos were cultured in PZM-3 for 6 days without activation treatment (EF group). Some of these embryos were additionally activated by electric stimulus (ES; EF+ES group) or a combination of ES and DMAP (EF+ES+D group) before culture. The reconstituted embryos were fixed 2.5h after fusion to evaluate the nuclear remodeling patterns. The nuclear remodeling pattern of NTs reconstituted with 44 h-matured recipients showed a tendency to form a pronucleus-like structure, while that of NTs reconstituted with 36 h-matured recipients showed a tendency to undergo a premature chromosome condensation (PCC) and form one set of chromatin clump. In EF+ES+D group, blastocyst development was significantly increased regardless of maturational age of recipient oocytes (P<0.05). The result indicates that additional activation treatment is necessary to induce the activation of embryos reconstituted with 36 h-matured recipients, and treatment with the combination of electrical stimuli and DMAP could enhance the blastocyst formation rate of porcine NTs reconstituted with both 36 h- and 44 h-matured recipient oocytes.

Animals↗
PubMed1989

In vitro fertilization and embryo transfer in the rhesus monkey.

Twenty-three rhesus monkeys were subjected to 9 days of ovarian hyperstimulation with sequential exposure to human follicle-stimulating hormone (hFSH) and then human luteinizing hormone (hLH) + hFSH. Six animals (26%) did not exhibit sustained, elevated levels of circulating estradiol, primarily due to the occurrence of a premature surge of endogenous LH (n = 4). Seventeen animals (74%) responded with supraphysiologic levels of circulating estradiol (peak value: means = 4480 pg/ml) and received human chorionic gonadotropin (hCG) on Day 10. Oocytes were collected 26 h later by aspiration of large antral follicles. Oocyte quantity (means = 18/animal) and quality (63% mature) were evaluated by in vitro fertilization (IVF), embryonic development, and embryo transfer to foster mothers. Modified conditions for the successful fertilization of oocytes used a Tyrode's augmented (TALP) medium supplemented with 0.3% bovine serum albumin (BSA). Oocytes were inseminated at the metaphase II stage with ejaculated, washed sperm (50 100 x 10(3)/ml) preexposed at ambient temperature to caffeine and dibutyryl cyclic adenosine 3'5'-monophosphate. Successful fertilization ranged from 26% to 75%. In one experiment, 5 of 11 embryos produced by IVF developed in vitro to hatched blastocysts. Embryo freezing employed a propanediol-based protocol and was applied to early cleavage-stage embryos with 100% (5 of 5) post-thaw survival. Two frozen-thawed embryos were transferred transtubally on 3 occasions into rhesus monkeys during the early luteal phase of spontaneous menstrual cycles. One pregnancy resulted, which proceeded normally to the unassisted delivery of a male offspring 170 days after the LH surge. We conclude that this sequential regimen of human gonadotropins provides a cohort of oocytes from rhesus monkeys that will complete meiotic maturation and fertilize in vitro, with embryonic development proceeding in vitro and in vivo. The production of putative antibodies to human gonadotropins, assessed by the presence of Protein A-precipitated hCG binding components in sera, limits the repeated use of monkeys in the hyperstimulation protocol. Nevertheless, this model system should facilitate further studies on oocyte maturation, fertilization, and early embryogenesis in primates.

Animals↗