Dose Of Follicle-stimulating Hormone Research Articles

Use of Triggers on in vitro Fertilization and Evaluation of Risk Factors for Sub-Optimal Maturation Rate.

To compare the oocyte maturation rate in the treatment of in vitro fertilization (IVF) in terms of the use of human chorionic gonadotropin (hCG), agonist gonadotropin-releasing hormone (GnRH) and dual trigger and to evaluate the associated risk factors for sub-optimal maturation rates. A retrospective cohort study with 856 women who underwent IVF. They performed oocyte retrieval and were classified into 3 groups (1 - hCG, 2 - GnRH agonist, 3 - dual trigger). The primary outcome was maturation rate per trigger, and the secondary outcomes were the pregnancy rate per oocyte retrieval and the correlations between low maturation rate as well as the clinical and treatment characteristics of women. The maturation rate was 77% in group 1; 76% in group 2, and 83% in group 3 (p = 0.003). Group 2 showed women with better ovarian reserve, greater number of oocytes collected, and more mature oocytes and embryos compared with the other groups (p < 0.001). The cumulative clinical pregnancy rate was no different between the groups (p = 0.755). Low ovarian reserve and low doses of follicle-stimulating hormone (FSH) administered during the stimulus were associated with a higher chance of null maturation rate. The oocyte maturation rates and IVF results were similar in all groups. Low ovarian reserve is associated with the worst treatment results.

Strategies for oocyte collection and in vitro embryo production in free-roaming bison herds.

The study was conducted to test the feasibility of protocols for field collection of cumulus–oocyte complexes (COC) for in vitro embryo production (IVP) in wild bison. The study was done with captive wood bison during the anovulatory season. In Experiment 1, the efficiency of transvaginal ultrasound-guided COC collection was compared between bison restrained in a squeeze chute without sedation vs in lateral recumbency after chemical immobilization using a dart gun (n = 8/group). In Experiment 2, a 2 × 2 design was used to examine the effects of superstimulation treatment [single dose of equine chorionic gonodotrophin (eCG) vs multiple doses of follicle stimulating hormone (FSH)] and method of drug administration (manual injection vs field darting) on COC collection and IVP. In Experiment 1, no difference was detected between chute-restrained vs chemically immobilized groups in the time required to complete COC collections, the number of follicles aspirated (11.5 ± 1.9 vs 9.3 ± 1.8; P = 0.4) or the COC recovery rate [COC recovered/follicle aspirated; 58/92 (63%) vs 44/69 (64%); P = 0.9]. In Experiment 2, no differences were detected between superstimulation treatments (eCG vs FSH). The total number of follicles available for aspiration did not differ between manual injection and field darting (23.9 ± 2.7 vs 21.6 ± 1.9; P = 0.4). Compared with the random start unstimulated group, the embryo production rate was higher [18/132 (14%) vs 53/189 (28%); P = 0.04] after wave synchronization and superstimulation. Results suggest that COC collection is equally feasible in a recumbent position after chemical immobilization as those bison restrained in a standing position in a hydraulic chute. Ovarian superstimulation with a single-dose eCG protocol is as effective as a multiple-dose FSH protocol, and field darting is as effective as chute-side administration of superstimulation treatments. The strategies in the present study are ready to be incorporated into field collections in free-roaming bison herds.

The Effect of Epidural Application of FSH on Superovulatory Response in Eastern Anatolian Red Cow

This study aimed to evaluate the superovulation response in Eastern Anatolian Red (EAR) cows treated with a single epidural injection or traditional administration of follicle stimulating hormone (FSH). Eight cows were randomly divided into two groups, as control (CG) and experiment (EG). Four replications were applied for each animal in the groups. A total of 16 studies were carried out in each group. In this study, 300 mg intramuscular (i.m) FSH injection was applied with decreasing doses twice daily for four days to the cows in the CG. (70:70, 50:50, 20:20, 10:10 mg). On the other hand, 300 mg FSH was administered as 150 mg epidural and 150 mg intramuscular injection to the cows in EG. The number of untransferable embryos and the total CLs were found statistically significant (p&amp;lt;0.05). However, the number of transferable embryos and unfertilized oocyte (UFO) were found statistically insignificant among the groups (p&amp;gt;0.05). According to the obtained data, it was observed that the epidural administration of FSH for superovulation in EAR cows did not provide sufficient follicle and oocyte development. In conclusion, different studies with different superovulation protocols and FSH doses in local breed EAR cows will be needed for further studies.

P–604 Effectiveness comparison of antral follicular count (AFC), follicular-output-rate (FORT), follicle-to-oocyte-index (FOI), oocyte-sensitivity-index (OSI), and follicular-sensitivity-idex (FSI) for predicting clinical pregnancy rates in IVF

Abstract Study question Which is better for predicting clinical pregnancy rate : AFC, FORT, FOI, FSI, or OSI? Summary answer Both AFC and OSI can be used to predict clinical pregnancy better than FORT, FOI or FSI. What is known already AFC, FORT, FOI, OSI, FSI can be used to predict clinical pregnancy, but no study compared which one is better Study design, size, duration Retrospective study using data from medical record (2016–2018) Subjects were patients underwent IVF cycle at Aster Clinic in Hasan Sadikin Hospital Bandung. Subjects divided into 2 groups: clinically pregnant that is visible gestational sac on ultrasound (n = 83) and not pregnant (n = 148). Inclusion criteria : antagonist protocols, &amp;lt;45 years, basal follicle stimulating hormone (FSH) ≤ 12 IU/L, ICSI fertilization method, and fresh transfer cycle. Participants/materials, setting, methods AFC categorized &amp;lt; 5 and ≥ 5 (poseidon) FORT=pre-ovulatory follicles(16–20 mm) x 100 divided by AFC(2–10 mm). FOI=oocytes obtained x 100 divided by AFC. OSI=oocytes obtained x 1000 divided by total FSH dose. FSI=pre-ovulatory follicles x 100,000 divided by (AFC x total FSH dose). FORT and FSI divided using percentil 33 and 67. OSI divided into 3 groups by cut-off 1.697/IU for poor-response and 10.07/IU for hyperresponse. FOI divided into 2 groups, ≤ 50% or &amp;gt; 50% Main results and the role of chance Group of AFC ≥ 5 had a significantly higher clinical pregnancy rate than the AFC &amp;lt; 5 group (39.49% vs. 16.67% ; p = 0.009). High and moderate OSI had higher clinical pregnancy rate than low OSI (66.37% vs. 37.72% vs. 25.45% ; p = 0.038). There is a significant negative correlation between OSI and age (–0.454) or total FSH dose (–0.594). There is a significant positive correlation between OSI and AFC (0.625), the number of follicles at trigger (0.792), and oocytes (0.923). There were no significant differences in clinical pregnancy rates between the FORT, FOI, and FSI groups. Limitations, reasons for caution Limitation Retrospective study using medical record data Ultrasound measurement was done by many reproductive gynecology specialist (not 1 person) --- observer bias. Wider implications of the findings: This study found no association between FORT, FOI, FSI on clinical pregnancy. Why? FORT, FSI, FOI use measurement number of follicles at trigger and antral follicle. Differences among observers in interpreting antral follicles and number of follicles at trigger, or inaccurate measurement. No FORT, FOI, and FSI cut off values from previous study. Trial registration number Not applicable

P–432 Do stage and grade of malignancy impact fertility preservation in breast cancer patients?

Abstract Study question Do stage and grade of breast cancer impact the number of retrieved mature oocytes during controlled ovarian stimulation for fertility preservation? Summary answer Stage and grade of breast cancer do not impact the number of retrieved mature oocytes. Higher grade breast cancer requires higher gonadotropin doses during stimulation. What is known already Cancer can impair ovarian response by unknown mechanisms. Some authors suggest that it could be detrimental on fertility because it elicits a catabolic state, increasing stress hormone levels. Some studies have also shown that ovarian response to controlled ovarian stimulation (COS) is, in some way, compromised in oncological patients. Little is known about the impact of different types of cancer on ovarian reserve, and specifically whether higher stage and grade could compromise egg retrieval during fertility preservation (FP) techniques. Study design, size, duration: Retrospective cohort study evaluating data of FP treatment cycles among women with breast cancer at the Oncofertility Unit of San Raffaele Hospital, Milan in the period from 2011 to 2019. Participants/materials, setting, methods Inclusion criteria were: breast cancer diagnosis; age 22–41; oocyte cryopreservation after stimulation with a random start GnRH-antagonist protocol. Patients receiving chemotherapy before FP were excluded. We compared outcomes between low-stage (stage I) and high-stage (stage II-III) patients and low-grade (G1-G2) and high-grade (G3) patients. Main study outcome was the total number of retrieved mature oocytes. Univariate analysis was performed by Mann-Whitney test, Kruskal-Wallis test and Fisher’s exact test. Multivariate analysis was performed by logistic regression. Main results and the role of chance 101 stimulation cycles were included. High-stage disease patients were significantly younger than low-stage. Median antral follicle count (AFC) was 12 in low-stage and 10 in high-stage (age-adjusted p = 0.92) and median anti-mullerian hormone (AMH) levels were 1.9 ug/L in low-stage and 1.8 ug/L in high-stage (age-adjusted p = 0.22). No significant difference in stimulation protocols and follicle-stimulating hormone (FSH) start and total dose could be detected between the 2 groups. Median number of vitrified oocytes was 7 in both groups (p = 0.75). No significant difference could be observed in median AFC (13 vs 10, p = 0.14) and AMH levels (2.1 vs 1.5, p = 0.88) between low-grade and high-grade disease patients. When adjusting for age, AFC was found to be significantly lower in high-grade disease patients (p = 0.03). Patients with high-grade tumors were stimulated with higher doses of FSH (age-adjusted p-value=0.03). Median number of vitrified oocytes was 6 in low-grade patients and 7 in high-grade (p = 0.35). In a multivariate model including age, cancer stage, cancer grade and molecular classification, the only significant factor found to be inversely associated with AFC was cancer grade (OR 3.6; 95% CI 0.7 – 6.5, p = 0.01), while only age was significantly associated with oocyte retrieval (OR 0.4; 95% CI 0.01 – 0.9, p = 0.04). Limitations, reasons for caution The main limitations of our study are its retrospective design and the small sample size. Wider implications of the findings: Fertility preservation counselling and ovarian stimulation protocols of breast cancer patients could be implemented with cancer grade. Trial registration number Not applicable

Effect of FSH and eCG on Alpaca (Vicugna pacos) Oocyte Maturation in vitro

This study evaluated alpaca oocytes that developed to metaphase II (MII), using different concentrations of follicle stimulating hormone (FSH) with and without equine chorionic gonadotropin (eCG) during in vitro maturation. Oocytes were obtained from ovaries of slaughtered alpacas. Oocytes were matured in vitro for 36h in TCM-99 and supplemented in groups with different doses of FSH: 0.5, 0.25, 0μg.mL-1 and eCG: 15, 5, 0IU.mL-1. Oocytes were stained with 2% lacmoid, and examined for their nuclear status. The parameter for comparisons between groups was the percentage of oocytes in MII. There was an interaction between FSH and eCG (P&lt;0.05). A higher percentage of oocytes in MII were obtained when using 0.5µg.mL-1 of FSH with any concentration of 15, 5 or 0IU.mL-1 of eCG [58.4±1.94% (n=78); 59.5±1.94% (n=85); 54.3±0.56% (n=81); P&lt;0.05]. A similar percentage was also found using 0.25µg.mL-1 of FSH plus 15IU.mL-1 of eCG [52.3±1.66% (n=86); P&gt;0.05] followed by 0.25µg.mL-1 of FSH with 5 or 0IU.mL-1 of eCG [47.3±1.94% (n=82); 45.3±1.94% (n=86); P&lt;0.05]. The lowest maturation percentages were found using any concentration of eCG without FSH [8.2±0.73% (n=84); 8.0±0.66% (n=74); 11.4±1.23% (n=71); P&lt;0.05]. In conclusion, the addition of eCG to the maturation medium would reduce the amount of FSH required in the in vitro maturation of alpaca oocytes.

Retrospective Monocentric Clinical Study on Male Infertility: Comparison between Two Different Therapeutic Schemes Using Follicle-Stimulating Hormone.

Follicle-stimulating hormone (FSH) is a therapeutic option in patients with idiopathic oligozoospermia and normal FSH serum levels. However, few studies have evaluated which dose of FSH is more effective. The aim of this study was to compare the clinical efficacy of the two most frequently used FSH treatment regimens: 75 IU daily vs. 150 IU three times a week. Patients were retrospectively assigned to two groups. The first group (n = 24) was prescribed highly purified FSH (hpFSH) 75 IU/daily (Group A), and the second group (n = 24) was prescribed hpFSH 150 IU three times a week (Group B) for three months. Before and after treatment, each patient underwent semen analysis, evaluation of the percentage of DNA-fragmented spermatozoa, assessment of testicular volume (by ultrasonography), and measurement of FSH and total testosterone (TT) serum levels. Treatment with hpFSH significantly improved conventional sperm parameters. In detail, sperm concentration increased significantly after treatment only in Group A, whereas total sperm count, percentage of spermatozoa with progressive motility, normal morphology, or alive improved significantly in both groups. Interestingly, the percentage of sperm DNA fragmentation decreased significantly in both groups after treatment with hpFSH. FSH serum levels were expectably higher at the end of the treatment than before hpFSH was administered to both groups. Remarkably, TT serum levels only increased significantly in Group A. Finally, testicular volume was significantly higher in Group A after treatment, while it did not change significantly compared to baseline in Group B. The percentage of FSH responders did not differ significantly between the two groups (8/24 vs. 6/24). The daily administration of hpFSH 75 IU seems more effective than using 150 IU three times a week. However, this therapeutic scheme implies a higher number of injections and slightly higher costs.

Effect of polymorphisms of MTHFR in controlled ovarian stimulation: a systematic review and meta-analysis.

Although several studies have reported a potential impact of methylenetetrahydrofolate reductase (MTHFR) polymorphisms on controlled ovarian stimulation (COS), the results remain controversial. The aim of the systematic review and meta-analysis was to evaluate the effect of MTHFR polymorphism on COS outcomes. PubMed, Web of Science, Embase, and Cochrane Central Register of Controlled Trials databases were searched up to December 2, 2020. COS clinical outcomes based on gene polymorphisms were included. Two reviewers independently extracted the data. The primary outcome was the number of oocytes retrieved. The secondary outcomes were the number of metaphase II (MII) oocytes, stimulation duration, basal follicle-stimulating hormone (FSH) level, FSH dosage, positive pregnancy test, ongoing pregnancy rate, clinical pregnancy rate, miscarriage rate, and live birth rate. Meta-analysis was performed using a fixed-effect model or random-effect model with Review Man 5.3.5. Mean difference (MD) with 95% confidence intervals (95%CIs) was calculated for continuous outcomes. The quality assessment of included studies was evaluated by using the Newcastle-Ottawa Scale. Eleven studies were included in the systematic review, and seven studies with 2015 participants were included in the meta-analysis. Basal FSH level was significantly lower in CC homozygotes than TT homozygotes (four studies, 867 participants, MD - 0.54, 95%CI - 0.85 to - 0.23, P = 0.0006; I2 = 0%) of MTHFR (rs1801133). FSH dose was significantly fewer in CC homozygotes compared with CT heterogeneous (three studies, 949 participants, MD - 75.78, 95%CI - 135.23 to - 16.33, P = 0.01; I2 = 32%) or CT/TT model (three studies, 1097 participants, MD - 80.18, 95%CI - 135.54 to - 24.81, P = 0.005; I2 = 42%). Differences in the oocytes retrieved and stimulation duration were insignificant. Gene variants on MTHFR (rs1801133) and MTHFR (rs1801131) were reported in ongoing pregnancy rate, clinical pregnancy rate, and live birth rate. Studies to date indicate that polymorphisms of MTHFR could influence basal FSH level and FSH dose. The results could be useful to promote clinical practice on COS protocols. Further studies are needed to evaluate the clinical relevance of the multigene combination on COS.

Curtailing the conventional dose of Folltropin-V for superstimulation and embryo recovery in Sahiwal cattle (Bos indicus)

High cost of the follicle stimulating hormone (FSH) impacted embryo transfer technology (ETT) work in India. The experiment evaluated effects of reduced (200 mg vs 400 mg) FSH (Folltropin-V) dose on superovulation and embryo recovery in Sahiwal cattle. Animals treated with conventional dose of Folltropin, i.e. 400 mg (G1, n=6) were compared with another group administered with half dose (i.e. 200 mg) of Folltropin (G2, n=6). Superstimulatory response (number of follicles &gt;8 mm) during estrus; unovulatory follicles, superovulatory response (one day prior to embryo flushing) using trans-rectal ultrasonography were recorded. Embryos were collected nonsurgically on day-7 post insemination and total recovered and transferable embryos were noted. Results revealed that number of follicles during estrus and subsequent ovulations were significantly higher in the G1 compared to G2 (30.0±1.37 and 26.3±0.99 vs. 20.3±0.21 and 17.7±0.42, respectively). Unovulatory follicles per animal were lower (2.6±0.21 vs. 3.7±0.49) in G2 than G1. The group receiving lower dose of Folltropin (G2) had significantly higher average embryo recovery compared to G1 (12.0±0.63 vs 8.7±0.76, respectively). Additionally, average number of transferable embryos were non-significantly higher in G2 (7.0±0.37) than G1 (5.7±0.88). The study indicated that lower dose of Folltropin-V (200 mg compared to 400 mg) is more effective in superovulation and embryo production in Sahiwal cows.

Discovery and Preclinical Development of Orally Active Small Molecules that Exhibit Highly Selective Follicle Stimulating Hormone Receptor Agonism.

An orally active follicle stimulating hormone receptor allosteric agonist would provide a preferred treatment for over 16 million infertile women of reproductive age in low complexity methods (ovulation induction-intrauterine insemination) or in high complexity methods (controlled ovarian stimulation-in vitro fertilization). We present two oral follicle stimulating hormone receptor allosteric agonist compounds that have the desired pharmacology, drug metabolism, pharmacokinetics, and safety profile for clinical use. These molecules provide a single agent suitable for ovulation induction-intrauterine insemination or controlled ovarian stimulation-in vitro fertilization that is more convenient for patients and achieves similar preclinical efficacy as rec-hFSH. TOP5668, TOP5300 were evaluated in vitro in Chinese hamster ovary cells transfected with individual glycoprotein receptors measuring cAMP (FSHR, LH/CGR, thyroid stimulating hormone receptor). TOP5668 was found to have solely follicle stimulating hormone receptor allosteric agonist activity while TOP5300 was found to have mixed follicle stimulating hormone receptor allosteric agonist and LHR-AA activity. Both compounds stimulated concentration-dependent increases in estradiol production from cultured rat granulosa cells in the presence or absence of low dose rec-hFSH, while only TOP5300 stimulated testosterone production from rat primary Leydig cells. In pooled human granulosa cells obtained from patients undergoing controlled ovarian stimulation-in vitro fertilization, TOP5300 stimulated 7-fold greater maximal estradiol response than rec-hFSH and TOP5668 was 10-fold more potent than TOP5300. Both TOP5300 and TOP5668 stimulated follicular development in immature rat to the same efficacy as recombinant follicle stimulating hormone. In mice treated with TOP5300, in the presence of low dose of follicle stimulating hormone, there were no differences in oocyte number, fertilization rate, and hatched blastocyst rate in mice with TOP5300 and low dose follicle stimulating hormone vs. reference proteins pregnant mare serum gonadotropin or high dose rec-hFSH. ADME/PK and safety profiles were favorable. In addition, there was no appreciable activity on thyroid hormones by TOP5300 in 14-days toxicological study in rat or dog. The selected lead compound, TOP5300 stimulated a more robust increase in estradiol production from granulosa-lutein cells from women with polycystic ovarian syndrome patient compared to rec-hFSH. Conclusions: Two novel oral FSHR allosteric agonist, TOP5668 and TOP5300, were found to mimic the biological activity of rec hFSH in preclinical studies. Both compounds led to folliculogenesis and superovulation in rat and mice. Specifically, TOP5300 led to a similar number of ovulated oocytes that fertilized and developed into hatched blastocysts in mice when compared to rec-hFSH. The safety profile demonstrated lack of toxicity.

Correlation of Anti-Mullerian Hormone Level and Antral Follicle Count with Oocyte Number in A Fixed-Dose Controlled Ovarian Hyperstimulation of Patients of In Vitro Fertilization Program.

BackgroundThis study was conducted to determine the correlation of anti-Mullerian hormone (AMH) level and antral follicle count (AFC) with oocyte count in women who had received controlled ovarian hyperstimulation in an in vitro fertilization (IVF) program.Materials and MethodsWe retrospectively gathered the data of 42 patients who underwent IVF during 2005-2017 at Aster Clinic in Dr. Hasan Sadikin Hospital and Bandung Fertility Center Limijati Hospital, Indonesia. Details of the subjects such as identity, characteristics, history of illness, history of previous therapy, levels of ovarian reserve markers examined (AFC and AMH), follicle-stimulating hormone (FSH) dose given, and number of oocytes produced were recorded.ResultsA significant positive correlation between AMH (P≤0.001, r=0.530), AFC (P≤0.001, r=0.687), and AMH- AFC combination (P≤0.001, r=0.652), and the number of oocytes was found at the FSH dose of 225 IU.ConclusionAFC and AMH are able to reliably predict ovarian response to FSH.

71 Effect of follicle-stimulating hormone dose and circulating progesterone before ovum pickup and invitro embryo production in pregnant Holstein heifers

Superstimulation with exogenous FSH before ovum pickup for invitro embryo production (IVP) improves embryo production. However, there is limited information on different FSH dose regimens and the effect of progesterone (P4) concentrations on IVP. Thus, the objective of the present study was to evaluate the effect of FSH dose and circulating P4 on ovarian superstimulation responses, before ovum pickup (OPU) on IVP in pregnant heifers. Pregnant Holstein heifers (n=47) 19.5±0.3 months of age with moderate body condition score (3.3±0.03; scale 1 to 5) were randomly assigned to 2 treatment groups, p-FSH (Folltropin-V, Vetoquinol) 160mg (FSH160) or p-FSH 300mg (FSH300), in a crossover design. Two sessions of OPU were performed on each animal at 48 and 62 days of gestation, with a washout interval between sessions of 14 days. Dominant follicle removal (DFR) was utilised to synchronize follicular wave emergence, and FSH treatments were initiated 36h later. Heifers in both groups received a total of 4 FSH injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3, and 32.0mg) or FSH300 (90.0, 80.0, 70.0, and 60.0mg). All heifers underwent OPU 40h after the last FSH treatment. Blood samples were collected for P4 quantitation at OPU and number of small (&amp;lt;6mm), medium (6–10mm), and large (&amp;gt;10mm) follicles were determined before OPU. Cumulus–oocyte complexes (COCs) recovered were processed and invitro embryo production was performed using standard procedures. Differences between treatment groups were evaluated using generalized linear mixed models, and data are presented as mean±s.e.m. (Table 1). Heifers treated with 300mg of p-FSH had a greater (P&amp;lt;0.05) number of medium-size follicles, total number of follicles, total COCs recovered, number of viable COCs, and number of blastocysts produced. In addition, heifers treated with 300mg of p-FSH had a greater cleavage rate (P=0.03). Circulating P4 was on average 10.1±0.4ng mL−1 and did not differ between gestation days (P=0.12). Furthermore, circulating P4 did not affect (P&amp;gt;0.10) any of the outcomes evaluated, nor was there an FSH dose by circulating P4 interaction (P&amp;gt;0.05). In conclusion, increasing the dose of p-FSH before OPU from 160 to 300mg resulted in an increase in the total number of follicles, number of oocytes, and number of blastocysts produced in pregnant donors without changes in blastocyst rate, regardless of progesterone concentrations at the time of OPU. Table 1. Ovum pickup and invitro embryo production in pregnant heifers treated with different doses of p-FSH Item FSH160 FSH300 P-value Small follicles (n) 5.0±0.6 3.5±0.4 0.02 Medium follicles (n) 14.4±0.8 18.3±0.9 0.0003 Large follicles (n) 2.9±0.3 2.5±0.3 0.28 Total follicles (n) 22.3±0.7 24.3±0.9 0.04 Total oocytes (n) 12.8±0.7 14.7±1.0 0.02 Recovery rate (%) 56.2±2.1 59.7±2.7 0.20 Viable oocytes (n) 11.4±0.7 13.3±1.0 0.02 Viable oocytes (%) 89.9±1.7 88.0±2.4 0.22 Cleavage rate (%) 63.8±4.3 71.5±3.7 0.03 Cleaved oocytes (n) 7.2±0.7 9.3±0.9 0.14 Blastocysts (n) 2.7±0.4 3.6±0.7 0.02 Blastocyst rate (%) 22.0±3.3 22.4±3.6 0.74

High doses of FSH induce autophagy in bovine ovarian granulosa cells via the AKT/mTOR pathway.

Follicle-stimulating hormone (FSH) plays a critical role in follicular growth and granulosa cell function; however, the mechanism by which the aggressive stimulation of FSH leads to poorer oocyte quality and embryo development potential is unclear. In this study, bovine ovarian granulosa cells (BGCs) were challenged with FSH doses (vehicle, 0.1, 1, 10 and 100ng/ml) to investigate the effects of FSH on BGCs. The results indicated that the relative viability of BGCs was significantly increased in cells challenged with 1ng/ml FSH, whereas the viability was significantly decreased with 100ng/ml FSH treatment. The mRNA abundance of FSHR, CYP19, StAR and BAX was significantly upregulated with 1, 10 and 100ng/ml of FSH, while the BCL-2 mRNA level was downregulated with higher concentrations of FSH (10 and 100ng/ml). Furthermore, BGC autophagy was detected in cells treated with 10 and 100ng/ml FSH by MDC staining, and the mRNA abundance of LC3, BECN1, BNIP3, ATG3 and ATG7 was upregulated with increasing FSH concentration. Meanwhile, the protein expression of LC3 was increased in cells treated with 10 and 100ng/ml FSH. 1 and 10ng/ml FSH significantly increased E2 production, whereas 10 and 100ng/ml FSH significantly increased P4 production. FSH significantly inhibited the phosphorylation of AKT in cells treated with higher concentrations (1, 10 and 100ng/ml), while activating mTOR phosphorylation at concentrations of 10 and 100ng/ml of FSH. In summary, we can conclude that higher doses of FSH (10 and 100ng/ml) induce BGC autophagy via the AKT/mTOR signalling pathway.

Negative impact of high doses of follicle-stimulating hormone during superovulation on the ovulatory follicle function in small ovarian reserve dairy heifers†.

When women with small ovarian reserves are subjected to assisted reproductive technologies, high doses of gonadotropins are linked to high oocyte and embryo wastage and low live birth rates. We hypothesized that excessive follicle-stimulating hormone (FSH) doses during superovulation are detrimental to ovulatory follicle function in individuals with a small ovarian reserve. To test this hypothesis, heifers with small ovarian reserves were injected twice daily for 4 days, beginning on Day 1 of the estrous cycle with 35, 70, 140, or 210IU doses of Folltropin-V (FSH). Each heifer (n=8) was superovulated using a Williams Latin Square Design. During each superovulation regimen, three prostaglandin F2α injections were given at 12-h interval, starting at the seventh FSH injection to regress the newly formed corpus luteum (CL). Human chorionic gonadotropin was injected 12h after the last (8th) FSH injection to induce ovulation. Daily ultrasonography and blood sampling were used to determine the number and size of follicles and corpora lutea, uterine thickness, and circulating concentrations of estradiol, progesterone, and anti-Müllerian hormone (AMH). The highest doses of FSH did not increase AMH, progesterone, number of ovulatory-size follicles, uterine thickness, or number of CL. However, estradiol production and ovulation rate were lower for heifers given high FSH doses compared to lower doses, indicating detrimental effects on ovulatory follicle function.

Effect of a dual trigger on oocyte maturation in young women with decreased ovarian reserve for the purpose of elective oocyte cryopreservation.

ObjectiveThe aim of this study was to determine whether co-administration of a gonadotropin-releasing hormone (GnRH) agonist and human chorionic gonadotropin (hCG) for final oocyte maturation improved mature oocyte cryopreservation outcomes in young women with decreased ovarian reserve (DOR) compared with hCG alone.MethodsBetween January 2016 and August 2019, controlled ovarian stimulation (COS) cycles in women (aged ≤35 years, anti-Müllerian hormone [AMH] <1.2 ng/mL) who underwent elective oocyte cryopreservation for fertility preservation were retrospectively analyzed.ResultsA total of 76 COS cycles were triggered with a GnRH agonist and hCG (the dual group) or hCG alone (the hCG group). The mean age and serum AMH levels were comparable between the two groups. The duration of stimulation, total dose of follicle-stimulating hormone used, and total number of oocytes retrieved were similar. However, the number of mature oocytes retrieved and the oocyte maturation rate were significantly higher in the dual group than in the hCG group (p=0.010 and p<0.001). After controlling for confounders, the dual-trigger method remained a significant factor related to the number of mature oocytes retrieved (p=0.016).ConclusionWe showed improved mature oocyte collection and maturation rate with the dual triggering of oocyte maturation in young women with DOR. A dual trigger appears to be more beneficial than hCG alone in terms of mature oocyte cryopreservation for young women with DOR.

FSH Stimulation with Short Withdrawal Improves Oocyte Competence in Italian Mediterranean Buffalo (Bubalus bubalis).

Simple SummaryThe high embryo production cost in buffalo, compared to cattle, is currently the major factor limiting the commercial application of ovum pick-up and in vitro embryo production technologies. This is mainly due to the lower number of follicles that develop during each estrous cycle and consequently oocytes recovered per session. In this work, we tested two different hormonal treatments based on commercial follicle-stimulating hormone (FSH) and progesterone (P4) to enhance the number of follicles and competent oocytes. Furthermore, in the second experiment, we tested three different coasting times, i.e., periods between the last FSH administration and ovarian aspiration, which is known be the key period for the final oocyte maturation and competence acquisition. The results, in terms of number of aspirated follicles, oocyte quality, blastocyst yield, and superior quality blastocyst yield, allow us to suggest the use of high doses of FSH (40 mg given six times every 12 h) combined with the shorter coasting time (28–32 h) as the ideal protocol for ovarian superstimulation in buffalo.The aim of this work was to evaluate the efficiency of different FSH doses and FSH coasting times before ovum pick-up (OPU) on follicular growth and oocyte competence in buffalo. Experiment 1 involved two different FSH treatments: 40 mg FSH given three (FSH3) or six (FSH6) times, 2 days after dominant follicle removal were tested, with OPU carried out after 40–44 h of coasting. In experiment 2, OPU was carried out after FSH6 protocol followed by 28–32 h (C1), 40–44 h (C2), or 64–68 h (C3) of coasting time. Cumulus oocyte complexes (COCs) were classified, in vitro matured, fertilized, and cultured. The results demonstrated that FSH6 increased the total number of follicles, the number and percentages of medium and large follicles, the number and the proportion of good quality oocytes, and the number of grade 1,2 and fast-developing blastocysts compared to the control. C3 decreased the percentage of good quality oocyte and blastocyst rates compared to C1 and C2. A higher percentage of fast blastocysts and average number of grade 1,2 blastocysts was observed in C1 compared to C3, with intermediate values found in C2. The improved efficiency in terms of blastocyst yields suggests the use of FSH6 + C1 protocol for ovarian superstimulation in buffalo.

Effect of low doses of FSH and season on the in vitro maturation, fertilization and embryo development of bovine oocytes

This study was aimed at determining the effect of follicle-stimulating hormone (FSH) and season on the in vitro maturation, fertilization and embryo development of bovine oocytes. Bovine ovaries obtained from a local slaughter house were transported to the laboratory within 2–3 h in a thermos flask containing antibiotic-supplemented physiological saline (0.9%) and at a fixed temperature of 30°C. Bovine oocytes collected in spring and autumn were incubated in culture media containing FSH at concentrations of 0.2 and 0.8 μg/ml. After maturation, oocytes were fertilized. Fertilized oocytes were incubated in CR1aa culture medium for 7 days at 38.5°C for in vitro development. The assessment made after the completion of the maturation process revealed that, for both FSH doses, the maturation rates obtained with the oocytes collected in spring were higher than those obtained with the oocytes collected in autumn. The incubation of the oocytes collected in autumn in culture media supplemented with 0.2 μg/ml of FSH resulted in a low level of oocyte maturation. After maturation, oocytes were subjected to fertilization. Fertilized oocytes were incubated in CR1aa culture medium for 7 days at 38.5°C for in vitro development. In both seasons, 0.8 μg/ml FSH application was higher than the maturation values obtained with 0.2 μg/ml FSH in terms of fertilization and embryo development rates. The study was repeated 9 times for each season. Although there was no significant difference between fertilizations and embryo development in the seasons, better results were obtained in spring season.

Identification of factors related to fertilization failure in in vitro fertilization-embryo transfer.

To investigate the possible factors relevant to fertilization failure in in vitro fertilization-embryo transfer (IVF-ET). The medical records of 4 205 infertile patients undergoing IVF-ET treatment at the Reproductive Medicine Center, Xiangya Hospital, Central South University from January 2016 to December 2017 were collected. The patients were divided into a complete fertilization failure group, a low fertilization rate group, and a control group based on fertilization rate. We examined the associations among the 3 groups in terms of female age, duration of infertility, duration of stimulation, gonadotropin (Gn) dosage, follicle-stimulating hormone (FSH) dosage, and total number of retrieved oocytes. According to theincidence factors, the patients were divided into a single female factor group, a single male factor group and a unisex factor group, and the correlation analysis of incidence factor among the 3 groups was performed. The patients were divided into a primary infertility and a secondary infertility in accordance with the type of infertility. We analyzed the correlation of infertility type among the three groups. Risk factors for complete fertilization failure and low fertilization rate in IVF-ET were obtained by stepwise multiple linear regression analysis. Primary infertility, long infertility duration, total number of retrieved oocytes, and unisex factor were associated with completefertilization failure and low fertilization rate in IVF-ET (P<0.05), but female age, duration of stimulation, FSH dosage as well as Gn dosage were not correlated with complete fertilization failure and low fertilization rate in IVF-ET (P>0.05). Stepwise multiple linear regression analysis showed that the incidence factor, type of infertility, and infertility duration were independent influential factors for complete fertilization failure and low fertilization rate. Complete fertilization failure and low fertilization rate in IVF-ET are related to duration of infertility, total number of retrieved oocytes, cause of onset, and type of infertility, but they are not relevant to female age, duration of stimulation, and Gn and FSH dosage.

Anti-Müllerian Hormone and Ovarian Reserve: Update on Assessing Ovarian Function.

ContextAnti-müllerian hormone (AMH) is produced by granulosa cells of small, growing follicles in the ovary. Serum AMH levels strongly correlate with the number of growing follicles, and therefore AMH has received increasing attention as a marker for ovarian reserve. This review summarizes recent findings and limitations in the application of serum AMH in ovarian reserve assessment.Evidence AcquisitionA PubMed search was conducted to find recent literature on the measurements and use of serum AMH as a marker for ovarian reserve.Evidence SynthesisSerum AMH levels are measured to assess the “functional ovarian reserve,” a term that is preferred over “ovarian reserve,” since AMH levels reflect the pool of growing follicles that potentially can ovulate. Serum AMH levels are used in individualized follicle-stimulating hormone dosing protocols and may predict the risk of poor response or ovarian hyperstimulation syndrome but has limited value in predicting ongoing pregnancy. Serum AMH levels are studied to predict natural or disease-related age of menopause. Studies show that the age-dependent decline rates of AMH vary among women. The generalized implementation of serum AMH measurement has also led to an increase in diagnostic assays, including automated assays. However, direct comparison of results remains problematic.ConclusionSerum AMH remains the preferred ovarian reserve marker. However, the lack of an international standard for AMH limits comparison between AMH assays. Furthermore, little is known about endogenous and exogenous factors that influence serum AMH levels, which limits proper interpretation of AMH values in a clinical setting.

Total follicle stimulating hormone dose is negatively correlated with live births in a donor/recipient model with fresh transfer: an analysis of 8,627 cycles from the Society for Assisted Reproductive Technology Registry

Oocyte donation has optimized our understanding of ovarian stimulation. Increasing the follicle-stimulating hormone (FSH) dose has been shown to adversely affect live birth rates in autologous cycles. Our objective is to assess whether this relationship holds true within the donor/recipient population. Retrospective cohort study. Not applicable. Data from 2014-2016 included 8,627 fresh donor cycles. None. Live birth, clinical pregnancy, and miscarriage rates. The mean donor age ± standard deviation (SD) was 25.8 ± 2.8 years. Donors underwent a median of 16 days (interquartile range [IQR] 12, 19) of stimulation with a median (IQR) total FSH dose and daily dose of 2,350.0 (1,800.0, 3,025.0) and 153.8 (113.2, 205.0) IU, respectively. The live birth rate was 56.7% per transfer. For every 500-unit increase in FSH dose, there was a 3% reduction in the odds of a live birth (odds ratio [OR] 0.97; 95% confidence interval 0.95, 0.99), and a 3% reduction in the odds of a clinical pregnancy (OR 0.97; 95% confidence interval 0.95, 0.99). Days of stimulation and average daily dose were not significantly associated with live birth or clinical pregnancy. No significant association was found between miscarriage rates and total FSH dose, days of stimulation, or average daily dose. This is a novel report of a negative association of total FSH dosage on fresh IVF live births, performed in the donor population to control for oocyte source and endometrial receptivity.