Induction Of Final Oocyte Maturation Research Articles

Personalization of IVF-ICSI workflow based on patient characteristics improves IVF laboratory outcomes and embryo ploidy by PGT-A

BackgroundIntracytoplasmic sperm injection (ICSI) has become a common method of fertilization in assisted reproduction worldwide. However, there are still gaps in knowledge of the ideal IVF-ICSI workflow including the optimal duration of time between induction of final oocyte maturation, oocyte denudation and ICSI. The aim of this study was to examine outcomes following different workflow protocols in IVF-ICSI procedures in blastocysts that have undergone undisturbed incubation and preimplantation genetic testing for aneuploidy (PGT-A) prior to transfer.MethodsRetrospective secondary analysis of 113 patients (179 IVF cycles, 713 embryos), all of whom have gone through IVF-ICSI and PGT-A using undisturbed culture. Predictive test variables were the length of time from: trigger to OPU, OPU to denudation, and denudation to ICSI. Outcome metrics assessed were: maturation, fertilization, blastulation and euploid rates. Generalized Estimated Equations Linear Model was used to examine the relationship between key elements of a given cycle and continuous outcomes and LOESS curves were used to determine the effect over time.ResultsIn a paired multi-regression analysis, where each patient served as its own control, delaying OPU in patients with unexplained infertility improved both maturation and blastulation rates (b = 29.7, p < 0.0001 and b = 9.1, p = 0.06, respectively). Longer incubation with cumulus cells (CCs) significantly correlated with improved ploidy rates among patients under 37, as well as among patients with unexplained infertility (r = 0.22 and 0.29, respectively), which was also evident in a multiple regression analysis (b = 6.73, p < 0.05), and in a paired analysis (b = 6.0, p < 0.05). Conversely, among patients with a leading infertility diagnosis of male factor, longer incubation of the denuded oocyte prior to ICSI resulted in a significantly higher euploid rate (b = 15.658, p < 0.0001).ConclusionsIn this study we have demonstrated that different IVF-ICSI workflows affect patients differently, depending on their primary infertility diagnosis. Thus, ideally, the IVF-ICSI workflow should be tailored to the individual patient based on the primary infertility diagnosis. This study contributes to our understanding surrounding the impact of IVF laboratory procedures and highlights the importance of not only tracking “classic” IVF outcomes (maturation, fertilization, blastulation rates), but highlights the importance that these procedures have on the ploidy of the embryo.

Kisspeptins stimulate the hypothalamus - pituitary - ovarian axis and induce final oocyte maturation and ovulation in female stinging catfish (Heteropneustes fossilis): Evidence from in vivo and in vitro studies

In the present study, effects of synthetic human Kisspeptin1 (hKiss1) and catfish Kisspeptin2 (cfKiss2) on the hypothalamic - pituitary - ovarian (HPO) axis, and induction of final oocyte maturation and ovulation were investigated in prespawning female stinging catfish (Heteropneustes fossilis) under in vivo and in vitro conditions. Gene expression was quantified by real time quantitative PCR and steroids were measured by specific enzyme-linked immunoassays. Intraperitoneal (ip) injections (1, 2, 3 ng/g body weight, BW) of synthetic hKiss1 and cfKiss2 stimulated hypothalamic, pituitary and ovarian gnrh1 and gnrh2 expression at 24 h and the effect was higher after the cfKiss2 treatment. In vitro incubation of hypothalamus, pituitary and ovary pieces with the Kiss peptides (5, 10, 20 nM) produced a similar effect. GPR54 (Kiss1 receptor) antagonist peptide234, when given ip (5, 10, 20 ng/g BW) or incubated in vitro (5, 10, 20 nM), inhibited the gnrh1 and gnrh2 expression at 24 h. The supplementation with hKiss1 or cfKiss2 restored the inhibition due to peptide234 in vivo and in vitro and the effect was higher in the cfKiss2 combination group. Both hKiss1 and cfKiss2 altered the expression of pituitary gonadotropin (Gth) subunit genes follicle-stimulating hormone β (fshβ), luteinizing hormone β (lhβ) and glycoprotein α (gpα) in vivo and in vitro. The expression of fshβ was more sensitive to the treatments than lhβ expression and the effect was greater in the cfKiss2 groups. Peptide234 in vivo and in vitro inhibited the expression of the Gth genes and the effect was reversed and restored in the hKiss1 and cfKiss2 combination groups. The Kiss peptide treatments in vivo or in vitro stimulated both plasma and ovarian levels of estradiol-17β, progesterone and 17,20β-dihydoxy-4-pregnen-3-one levels. The peptide234 treatment inhibited, or elicited a decreasing trend on the steroid levels both in vivo and in vitro, and the inhibition was reversed by the hKiss1 and cfKiss2 combination treatments. Incubation of post vitellogenic follicles with hKiss1 or cfKiss2 stimulated germinal vesicle breakdown (GVBD) and ovulation. The inhibition due to peptide234 was reversed in the combination groups. Ovulation was not elicited or unaffected in the peptide234 treated groups. The data show that the Kiss peptides act downstream the HPO axis to stimulate oocyte maturation and ovulation, and cfKiss2 peptide is functionally more effective than hKiss1.

Role of GnRH and GnIH in paracrine/autocrine control of final oocyte maturation

The control of oocyte growth and its final maturation is multifactorial and involves a number of hypothalamic, hypophyseal, and peripheral hormones. In this study, we investigated the direct actions of the gonadotropin-releasing hormone (GnRH) and the gonadotropin-inhibitory hormone (GnIH), which are expressed in the ovarian follicles, on final oocyte maturation in zebrafish, in vitro. Our study demonstrates the expression of GnRH and GnIH in the ovarian follicles of zebrafish (Danio rerio) at different stages of development and provides information on the direct action of these hormones on final oocyte maturation. Treatment with both GnRH and GnIH peptides stimulated the germinal vesicle breakdown (GVBD) of the late-vitellogenic oocyte. Both the GnRH and GnIH treatments showed no significant change in the caspase-3 activity of pre-vitellogenic and mid-vitellogenic oocytes, while they displayed different responses in the late-vitellogenic follicles. The GnRH treatment increased caspase-3 activity, whereas the GnIH reduced caspase-3 activity in the late-vitellogenic follicles. We also investigated the effects of GnRH and GnIH on the hCG-induced resumption of meiosis and caspase activity in vitro. GnRH and GnIH were found to have a similar effect on the hCG-induced resumption of meiosis, while they showed the opposite effect on caspase-3 activity. Furthermore, we investigated the effects of concomitant treatment of GnRH and GnIH peptides with hCG. The results demonstrated that the presence of both GnRH3 and GnIH are necessary for the normal induction of final oocyte maturation by gonadotropins. The findings support the hypothesis that GnIH and GnRH peptides produced in the ovary are part of a complex multifactorial regulatory system that controls zebrafish final oocyte maturation in paracrine/autocrine manner working in concert with gonadotropin hormones.

Out-of-season artificial reproduction of common dace (Leuciscus leuciscus L.) under controlled conditions

This study focused on the artificial reproduction of common dace Leuciscus leuciscus (L.) outside the reproductive season. The results indicate there is the possibility of initiating the reproducion in individuals of this before the natural spawning season. There could be induction of spermiation by altering the environmental conditions. For females, hormonal injections were necessary for induction of final oocyte maturation and ovulation. Generally, there were high percentages of spermiation (100%) and ovulation (87%–100%) as well as low mortality (0%–11% and 7%–13% for males and females, respectively) among the induced spawners. The greatest embryo survival occurred when Ovaprim (84.4%) or hCG (89.6%) was administered, although the latency time using hCG was at least twice as long compared to when other spawning agents were used (84–92 hrs and 30–44 hrs). The results from present study could be applicable for optimization of breeding stock numbers in aquaculture enterprises and in providing insights for conservation of L. leuciscus endangered populations.

An in vitro study on noradrenergic modulation of final oocyte maturation in the catfish Heteropneustes fossilis

This study was aimed to elucidate in vitro the effects of endogenous catecholamines: dopamine (DA), noradrenaline (NA) and adrenaline (A), and the β-adrenergic blocker propranolol on induction of final oocyte maturation (FOM) in the catfish Heteropneustes fossilis. With this aim, post vitellogenic follicles from sexually mature gravid female catfish were incubated with each of DA, NA and A in a concentration range of 5–250 μM, and propranolol in a concentration range of 1–200 μg/mL at time points varying from 0 to 30 h). Translucent follicles without germinal vesicle (GV) and opaque follicles with GV were scored separately for the calculation of percentage germinal vesicle breakdown (GVBD), an index of FOM. Data were analyzed by one-way ANOVA and were considered statistically significant when P values were less than 0.05. The analysis of the data showed that the incubation with NA only stimulated GVBD in a concentration - and time-dependent manner. Though the incubation with propranolol decreased total follicular cAMP level significantly at and above 10 μg/mL concentrations, a significant effect of the GVBD increase was noticed at 50 μg/mL or higher. However, the 10 μg/mL concentration of propranolol was effective to inhibit the NA-induced GVBD significantly albeit at a low level (39%). The present study suggests that final oocyte maturation is modulated by NA through a β-adrenergic mechanism, implicating a neural control of oocyte maturation and ovulation in teleosts.

Hormonal induction of ovulation using Ovaprim™ [(D-Arg6, Pro9NEt)-sGnRH + domperidone] and its impact on embryonic development of wild-caught Longspine scraper, Capoeta trutta (Heckel, 1843)

Knowledge of gamete quality is a prerequisite for developing techniques to fertilize eggs and rear offspring for hatchery production. Our objective was to develop assisted reproductive techniques, via hormonal induction of final oocyte maturation (FOM), for Longspine scraper, Capoeta trutta. Fish were administered injections of salmon gonadotropin-releasing hormone analogue containing anti-dopaminergic drug (Ovaprim™) or saline (control). Effects of Ovaprim on induction of ovulation, gamete quality, embryonic development, and larval survival were later examined with serum steroid hormone levels and ovarian histology. The saline group failed to spawn, whereas Ovaprim accelerated FOM and induced spawning. Fish treated with Ovaprim showed an increase in gonadosomatic index, egg diameter, and wet weight relative to controls. Average absolute fecundity, relative fecundity, fertilization, and hatching rates were 8823 eggs/spawn, 53 eggs/g body weight, 95%, and 91%, respectively. Serum 17α,20β-dihydroxy-4-pregnen-3-one (DHP) levels were significantly enhanced by ∼4-fold in Ovaprim-treated fish compared to the saline-injected fish, while 17β-estradiol levels declined upon FOM in hormone treated fish. Embryonic development closely resembled the teleost scheme, despite variations in timing. Larval survival at 6 and 12days post-hatch were 98% and 95%, respectively. Results suggest that Ovaprim is efficient for inducing spawning in C. trutta for stock enhancement or hatchery purposes.

Reproductive outcomes after a single dose of gonadotropin-releasing hormone agonist compared with human chorionic gonadotropin for the induction of final oocyte maturation in hyper-responder women aged 35–40 years

To investigate the reproductive outcomes after the use of GnRH agonist (GnRHa) compared with hCG for the induction of final oocyte maturation in GnRH antagonist cycles performed in hyper-responder women aged 35-40years. Retrospective study. Academic fertility center. Two hundred seventy-two hyper-responder women aged 35-40years who underwent controlled ovarian stimulation under GnRH antagonist suppression were included. Final oocyte maturation was performed with GnRHa (n = 168) or hCG (n = 104). Embryos were cryopreserved at the blastocyst stage and transferred in subsequent warming cycles (n = 542). Subjects were included in the analysis until live birth was achieved, after which they were excluded from further analysis. None. Cumulative live birth rate. Subjects in the GnRHa group achieved a higher number of oocytes (22 vs. 21) and a higher number of mature oocytes (16 vs. 14). The number of cryopreserved blastocysts (median of five blastocysts in both groups) was similar. Women in the hCG group needed a lower number of warming cycles to achieve live birth (1.32 vs. 2.12), had higher embryo implantation rates (48% vs. 39%), and the proportion of embryos transferred until live birth was lower (33% vs. 57%). The cumulative live birth rate was similar between the groups (48.15% vs. 48%). Although the cumulative live birth rate is similar, a single dose of GnRHa possibly results in suboptimal oocyte and embryo competence, as manifested by decreased embryo implantation rates and increased time needed to achieve live birth.

Evaluation of Ovaprim and human chorionic gonadotropin doses on spawning induction and egg and larval quality of pinfish, Lagodon rhomboides

The use of exogenous hormones to induce final oocyte maturation (FOM), ovulation, and spawning has become commonplace in the reproductive protocols for many species. Dose efficacy can be highly variable among species and factors such as temperature and stress may further obfuscate the hormone's perceived efficacy. The pinfish, Lagodon rhomboides, is a marine baitfish species with great potential for commercial development. This species readily undergoes vitellogenesis in captivity, however progression through FOM is unreliable. Therefore, the objectives of this investigation were to assess the effects of various doses of Ovaprim® (sGnRHa+domperidone) and human chorionic gonadotropin (HCG) on ovulation and spawning in pinfish.Two experiments were conducted to determine efficacious doses of Ovaprim and HCG in pinfish. Ovaprim dosages investigated were 0.25, 0.50, 1.00, and 2.00mL/kg, injected into the dorsal musculature of female pinfish. Male pinfish received one half the dosage (0.125, 0.25, 0.50, and 1.00mL/kg) administered to females to ensure spermiation. Experimental HCG doses of 500, 1000, 2000, and 4000IU/kg were administered to female pinfish with male pinfish once again receiving half the dosage (250, 500, 1000, and 2000IU/kg). Spawned eggs were collected over a 72hour period post injection and enumerated and assessed for fertilization, hatching percentage, and survival to first feeding. Additionally, eggs and larvae were photographed and a suite of morphological parameters was evaluated.Doses of 0.25 and 0.50mL/kg Ovaprim were efficient at induction of FOM and spawning in pinfish. Superior fecundity, fertilization and 3day post hatch (DPH) survival rates, as well as larger 3 DPH larvae support lower Ovaprim doses as the preferred choice for use with this species. Conversely, the 4000IU/kg HCG dose performed the best of all HCG doses evaluated eliciting the greatest spawning frequency from female pinfish.

Low-dose hCG supplementation after GnRH agonist triggering: don't be too quick on the trigger

Induction of final oocyte maturation using a GnRH agonist was first described in the early nineties, though with the advent of the GnRH antagonist era it has gained ever increasing attention some two decades later (Humaidan et al., 2011). The primary motive for substituting the ‘classical’ hCG trigger is the complete elimination of clinically significant ovarian hyperstimulation syndrome (OHSS). When used alone (or with steroid-only luteal support) GnRH agonist triggering is undoubtedly themost efficient and powerfulwayof (secondary)OHSSprevention. So far there is no confirmedOHSS case in the literature that proves the opposite (Kol and Humaidan, 2012). This prevention occurs due to a massive, rapid and irreversible luteolysis leading to a practical, total elimination of clinically significant moderate/severeOHSS (Kol, 2004). Consequently when fresh embryo transfer is not planned this strategy has established itself as the first choice option in oocyte donor/fertility preservation cycles and before total freezing of oocytes or embryos (Bodri et al., 2010; Griesinger et al., 2011). However, if the primary goal is performing fresh embryo transfer then the challenge is to save the severely impaired luteal phase without increasing the risk of early/late-onset OHSS. After the discouraging results of initial trials only a few dedicated research groups have made further efforts to undertake research in this area (Humaidan et al., 2006; Engmann et al., 2008; Castillo et al., 2010; Papanikolaou et al., 2011b). Humaidan et al. developed the concept of a modified luteal support aimed at rescuing the impaired luteal phase by administering small boluses of luteotropic substances. This hypothesis was first tested in pilot studies, both in normal and high responders (Humaidan et al., 2006 Humaidan, 2009) and later in a randomized clinical trial (Humaidan et al., 2010). The first randomized trial (mainly involving normal responders) suggested that the reproductive outcome can be improved to the level of control patients with a single, low-dose (1500 IU) hCG bolus administered at the time of oocyte retrieval, without increasing the risk of OHSS (Humaidan et al., 2010). These encouraging results were also reproduced in a retrospective study by an Australian group (Radesic and Tremellen, 2011). Although Radesic and Tremellen used the aforementioned strategy with a fairly high OHSS risk population (without a pre-established upper limit of ovarian response) they were cautious enough to only perform mandatory single embryo transfer to reduce the risk of any late-onset OHSS. In the current issue of Human Reproduction the case series by Seyhan et al., report the combined experience from two geographically distant centres in Canada and Turkey with IVF patients at high risk of OHSS, who received the low-dose hCG rescue protocol. Despite using co-interventions (such as coasting and cabergoline administration) the authors reported a worryingly high rate (22%) of early onset severe OHSS requiring hospitalization. Although the presented retrospective case series is fairly small, the authors felt that it was important to highlight that early onset OHSS can still occur after applying the low-dose hCG rescue protocol. At first sight their findings are in blunt disagreement with most previously published studies that have also used low-dose hCG luteal support (single or multiple-bolus) following GnRH agonist trigger (Table I). However, at closer look notable differences can be observed. Among the four studies that were conducted in high OHSS risk patients Seyhan et al.’s 23 patients represent the highest ovarian response category with extremes of 49 and 65 oocytes retrieved. Moreover, although in one of the participating centres total freeze was considered in 33% of the cases, in the other fresh transfer was performed only transferring two to three embryos. Taken altogether these factors considerably influence the risk of early and late-onset OHSS. Although it was based on a very limited number of cases (5 versus 18) Seyhan et al. also suggested a threshold (≥18 follicles) based on the number of intermediate-sized (10–14 mm) follicles above which embryo/oocyte freezing is strongly recommended. This threshold (which was the only significant finding in this series) was based on the number of intermediate follicles and might be less practical for everyday clinical use than the total follicular count (Papanikolaou et al., 2006). Interestingly, in the design of a more recent randomized trial by Humaidan et al. on low-dose hCG support, patients with .25 follicles .11 mm were judged to be at too high risk and excluded from participation (www.clinicaltrials.gov: NCT00627406).

Effect of different commercial spawning agents and thermal regime on the effectiveness of pikeperch, Sander lucioperca (L.), reproduction under controlled conditions

Pikeperch, Sander lucioperca (L.), has been identified as one of the most perspective candidates for diversification of freshwater aquaculture. However, some aspects of production are still being developed, and controlled reproduction is one of the bottlenecks. The aim of the present study was to compare the effectiveness of different commercial spawning agents in the induction of final oocyte maturation (FOM) and ovulation in wild spawners. Within the study, four spawning agents [human chorionic gonadotropin (hCG), mixed human and horse gonadotropin (PG-600), carp pituitary (CPH) and mammalian GnRH analogue combined with metoclopramide (Ovopel)] in different thermal regimes (13 and 15 °C) were tested. In both thermal regimes, the highest (P < 0.05) ovulation rate among the treatment groups was obtained after stimulation with hCG (100 % in both cases). Latency time was the shortest in groups where CPH was used (2–3 and 3–4 days for 15 and 13 °C) and was similar in the remaining groups (3–4 and 4–5 days for 15 and 13 °C, respectively). Embryo survival was the highest in groups treated with hCG (78.9 and 81.3 % at hatching stage for 15 and 13 °C, respectively). Hormonal stimulation did not significantly affect spermiation rate or spermatozoa motility (P > 0.05). Based on the obtained results, hCG can be recommended for induction of FOM and ovulation in pikeperch. In addition, the thermal regime within the tested range seemed to have no effect on the reproduction outcome, and the application of lower temperature only prolonged the time of ovulation.

Induction of Final Oocyte Maturation and Ovulation in Jack Mackerel, Trachurus japonicus, Temporarily Reared in Captivity

Induction of Final Oocyte Maturation and Ovulation in Jack Mackerel, Trachurus japonicus, Temporarily Reared in Captivity

While we are waiting for NFOG 2012 and summer, enjoy this issue of AOGS

While we are waiting for NFOG 2012 and summer, enjoy this issue of AOGS

Comparing the Efficacy of Urinary hCG vs Recombinant hCG for Final Maturation of Oocyte in GnRH Antagonist IVF/ICSI Cycle

ABSTRACTAims and objectivesTo compare the efficacy of urinary human chorionic gonadotropin (uhCG)vsrecombinant human chorionic gonadotropin (rhCG) for triggering ovulation in gonadotropinreleasing hormone (GnRH) antagonistin vitrofertilization (IVF) cycle.Materials and methodsTwo hundred patients who underwent GnRH antagonist IVF/ICSI cycles were analyzed, 100 of them received recombinant hCG 250 μg (injection overtrelle) and another group of 100 received urinary hCG (injection ovutrig) either 5000 IU or 10,000 IU for final oocyte maturation and ovulation trigger. The primary outcome measured was total oocytes retrieved and the secondary outcome measured 09 was the maturity rate fertilization rate, cleavage rate and clinical pregnancy rates were also compared.ResultsAge, day 2 follicle stimulating hormone (FSH) and total dose of rFSH were comparable between the groups. There were no significant differences between the groups in terms of the mean number of oocytes retrieved per follicle (uhCG; 12.5, rhCG; 12.09, p-value-0.6698). Similarly, there was statistically no difference in any of the other parameters studied between the two groups. That is number of mature oocytes, number of fertilized oocytes and number of cleaved oocytes, as the p-value was not significant. The clinical pregnancy rate was somewhat higher in the uhCG group but was not statistically significant. Both treatments were well tolerated and there was no significant side effects for either drug.ConclusionThere is no difference in clinical outcomes between urinary and recombinant hCG for induction of final oocyte maturation. Additional factors, including the cost, drug availability and ease of administration, should be considered, when choosing gonadotropin type.How to cite this articleSidhmalswamy GA, Srinivas MS, Dipika K, Anu K, Rao KA, Mekhala D. Comparing the Efficacy of Urinary hCG vs Recombinant hCG for Final Maturation of Oocyte in GnRH Antagonist IVF/ICSI Cycle. Int J Infertility Fetal Med 2012;3(3):92-96.

Fusion of lipid droplets in Japanese eel oocytes: Stage classification and its use as a biomarker for induction of final oocyte maturation and ovulation

The quality of eggs obtained from maturation-induced Japanese eel, Anguilla japonica, is unstable. One of the causes for low quality eggs is that females are artificially induced to ovulate at an inappropriate maturational status. Here we investigated the feasibility to use the morphological changes of lipid droplets in oocytes as a biomarker to show the optimum timing for injections of salmon pituitary extract (SPE) for priming and maturation-inducing steroid (MIS) for final oocyte maturation and ovulation. Various sizes of oocytes (700 to 1000 μm) were collected by cannulation from maturation-induced female eels during five days until ovulation. Morphology of the lipid droplets in these oocytes were classified into 10 stages mainly on the basis of their diameter, which increased with the progress of maturation as the droplets fused together. To assess the maturational status of each female, the lipid droplet stage was determined for 10 oocytes randomly chosen among the most advanced mode of developing oocytes and the median value was defined as the stage for the individual. Relationship between the lipid droplet stage at induction of ovulation and hatchability of the ovulated eggs was examined in 23 females, in which the hatching rate markedly fluctuated. Stages of females from which good quality eggs were obtained converged around stage 4 (five largest droplets, 55 to 70 μm) at priming with SPE, stage 6 (90 to 110 μm) at injection with MIS, and stage 7 (110 to 130 μm) at ovulation, while those of females from which low quality eggs were obtained showed distributions over broader ranges. Furthermore, 466 females were induced to mature and the lipid droplet stage at induction of ovulation was examined in 13 females from which high quality eggs (hatching rate >80%) were obtained. Most of the 13 females were at stage 3 (40 to 55 μm) or 4 when primed with SPE, at stage 6 when injected with MIS, and at stage 7 when they ovulated. These results suggest that the optimum stages for SPE priming, MIS injection, and ovulation are stages 3 to 4, 6, and 7, respectively, though the optimum stage for priming will be variable if the temperature and/or the time between priming and MIS injection is varied. We conclude that the lipid droplet stage can be used as a sensitive biomarker to show the optimum timing for inducing ovulation by hormonal treatments.

Exploring two different doses of GnRH agonist for the induction of final oocyte maturation in GnRH antagonist-treated oocyte donor cycles: a retrospective comparison

Ovarian stimulation with the GnRH antagonist protocol is a well established option in oocyte donor program. The induction of final oocyte maturation can be performed with a GnRH agonist instead of hCG. Nonetheless the type of agonist used or its dose are not well defined and further tailoring might still be needed.

Oocyte vitrification: influence of operator and learning time on survival and development parameters

The threat of severe ovarian hyperstimulation syndrome (OHSS) and the increase in discomfort for the patient has limited the feasibility of maximizing the oocyte yield per treatment cycle. A gonadotrophin-releasing hormone (GnRH) antagonist protocol with agonist triggering and vitrification of all 2PN oocytes can eliminate the risk of OHSS. This prospective, single-centre, cohort study in 30 good-responder IVF patients ⩽36 years reports the feasibility of arbitrarily intensifying stimulation in a GnRH antagonist protocol in terms of tolerability, safety and efficacy. Ovarian stimulation was performed with 225–375 IU FSH, induction of final oocyte maturation with 0.2 mg GnRH agonist followed by vitrification of all 2 pronuclear (2PN) oocytes and repetitive vitrified–warmed embryo transfer cycles. Main outcomes were severe OHSS incidence, tolerability, assessed by a questionnaire, and cumulative live birth rate. On average, 17 oocytes were retrieved (range 4–42) and 8.4 oocytes at the 2PN stage were cryopreserved (range 3–22). No case of severe OHSS was observed (0%, 95 CI 0–11.4%). Tolerability was good. The cumulative live birth rate per patient undergoing at least one vitrified–warmed embryo transfer was 26.9% (7/26, 95% CI 13.7–46.1%). This approach can be explored in future larger-sized controlled studies.The more oocytes that are available after ovarian stimulation, the more often a patient can undergo an embryo transfer when frozen–thawed cycles using cryopreserved embryos are taken into account. However, the feasibility of retrieving a large number of oocytes has so far been hampered by the risk of ovarian hyperstimulation syndrome (OHSS). An approach combining a gonadotrophin-releasing hormone (GnRH) antagonist protocol stimulation with agonist triggering of final oocyte maturation and vitrification of all 2 pronuclear oocytes eliminates the risk of OHSS. Herein we report the feasibility of arbitrarily intensifying stimulation in a GnRH antagonist protocol in terms of tolerability, safety and efficacy. It was found in a small study on young patients that harvesting a large number of oocytes (on average 17) is not associated with poor tolerability or an increased risk of OHSS. The live birth rate after vitrified–thawed embryo transfers was 26.9%. Thus, this approach can be studied in larger-sized controlled trials.

Triggering final oocyte maturation with reduced doses of hCG in IVF/ICSI: a prospective, randomized and controlled study

ObjectiveTo compare the effectiveness of urinary human chorionic gonadotropin (u-hCG) at reduced doses of 4000IU and 6000IU in inducing final oocyte maturation during in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles. Study design164 patients with an indication for IVF or ICSI recruited in this randomized, single-blinded and controlled study in IVF clinic at the Sun Yat-sen Memorial Hospital. Patients were prospectively randomized to receive 4000IU (Group A, n=83) and 6000IU (Group B, n=81) of hCG for triggering final oocyte maturation. Number or percentage of mature oocytes retrieved per patient, fertilization rates, pregnancy rates were the main outcome measures. ResultsNo evidence of statistically significant difference in the number or proportion of mature oocytes retrieved was observed in both groups. The lower fertilization rate and significantly lower clinical pregnancy rate were observed in Group A. The ovarian hyperstimulation syndrome (OHSS) rates in both groups were also similar. In the subgroup of BMI<20kg/m2, fertilization rate were significantly higher in the administration group of hCG at the dose of 6000IU when compared with the dose of 4000IU (82.40% vs. 70.92%, P=0.017); in contrast, no significant difference in clinical pregnancy rates was observed in both groups. In the subgroup of BMI 20–25kg/m2, clinical pregnancy rates were significantly higher in patients treated with hCG at dose of 6000IU than patients treated with hCG at dose of 4000IU (65.3% vs. 35.0%, P=0.004); however, no significant difference in fertilization rates was observed. ConclusionBoth doses of u-hCG revealed an equal effect on the induction of final oocyte maturation in the patients with moderate or high ovarian response; however, the reduced dose of hCG could result in an obvious impact on clinical pregnancy rates and did not exhibit an obvious effect on OHSS rates.

Vasotocin induces final oocyte maturation and ovulation through the production of a maturation-inducing steroid in the catfish Heteropneustes fossilis

The study reports for the first time vasotocin (VT) induction of final oocyte maturation and ovulation through the production of the maturation-inducing steroid 17, 20β-dihydroxy-4-pregnen-3-one (MIS, 17, 20β-DP). Post-vitellogenic follicles of the catfish Heteropneustes fossilis were incubated with different concentrations of VT (1, 10, 100 and 1000 nM) for different time periods. Germinal vesicle breakdown [GVBD, as a marker of final oocyte maturation (FOM)] and ovulation were scored. In another series of experiments, the follicles were incubated with VT alone or in combination with VT receptor (V 1 and V 2) antagonists, and GVBD and ovulation were increased with progesterone, 17-hydroxy-4-pregnene-3, 20-dione (17-P) and 17, 20β-DP levels. VT stimulated both GVBD and ovulation in a concentration and time-dependent manner, and the responses were inhibited to varying degrees in groups incubated with the VT receptor antagonists. The V 1 antagonist inhibited the responses by 2- to 3-fold and more than the V 2 antagonist, and the combination was more potent than the separate incubation. Progestins increased time-dependently in the VT groups and the fold increase was greater for the MIS. The VT-induced steroid stimulation was significantly inhibited to near the control levels in co-incubations with both V 1 and V 2 receptor antagonists, in the order 17, 20β-DP > 17-P > P 4. The inhibition by the V 1 receptor antagonist was greater than that with the V 2 blocker, and followed the same order of inhibition described above. The results suggest that VT induces FOM and ovulation mainly through the V 1 receptors.

Ovarian steroidogenesis and the role of sex steroid hormones on ovarian growth and maturation of the Japanese eel

Three sex steroid hormones, estradiol-17β (E2), 11-ketotestosterone (11-KT), and 17α,20β-dihydroxy-4-pregnen-3-one (DHP), are well established as primary estrogen, androgen, and progestin, respectively, in teleost fish. Japanese eel, Anguilla japonica, would be a suitable candidate to study ovarian steroid physiology of fish because the ovarian growth and steroidogenesis is dormant under laboratory condition but can be induced by administration of exogenous gonadotropic reagents. In this review, we summarized our work on the function and production of sex steroid hormones in the ovary of the Japanese eel during ovarian growth and oocyte maturation artificially induced by treatment with extract of salmon pituitary. In vitro and in vivo assays suggest that 11-KT and E2 play primary roles in previtellogenic and vitellogenic growth of oocytes, respectively, whereas DHP is essential for induction of final oocyte maturation. We also reviewed the correlation between ovarian steroidogenesis to produce these sex steroid hormones, serum titers and gene expression.

Follicle-stimulating hormone administered at the time of human chorionic gonadotropin trigger improves oocyte developmental competence in in vitro fertilization cycles: a randomized, double-blind, placebo-controlled trial

To determine whether an additional follicle-stimulating hormone (FSH) bolus administered at the time of the human chorionic gonadotropin (hCG) trigger can improve the developmental competence of the oocyte. Randomized, double-blind, placebo-controlled, clinical trial. Academic medical center. Women undergoing a long agonist suppression in vitro fertilization (IVF) protocol for treatment of infertility. FSH bolus at time of hCG trigger versus placebo. Primary outcome; fertilization; secondary outcomes: oocyte recovery, implantation rate, and clinical and ongoing pregnancy/live birth rates. A total of 188 women (mean age: 36.2 years; range: 25 to 40 years) were randomized. Fertilization (2PN/#oocyte) was statistically significantly improved in the treatment arm (63% vs. 55%) as was the likelihood of oocyte recovery (70% vs. 57%). There was no statistically significant difference in clinical pregnancy rate (56.8% vs. 46.2%) or ongoing/live birth rate (51.6% vs. 43.0%). Improvements in IVF success rates have largely been due to optimization of embryo culture and stimulation protocols; less attention has been directed toward methods to improve induction of final oocyte maturation. This was the first randomized, double-blind, placebo-controlled trial to modify the ovulation trigger to improve oocyte competence, as demonstrated by the statistically significant improvement in fertilization.