Analysis of maturation dynamics and developmental competence of in vitro matured oocytes under time-lapse monitoring

Background To improve the developmental competence of in vitro cultured oocytes, extensive literature focused on maturation rate improvement with different additives in culture medium, while studies investigating the maturation dynamics of oocytes during in vitro maturation (IVM) and the influencing factors on oocyte viability are scarce. Methods The study involved a retrospective observation by time-lapse monitoring of the IVM process of 157 donated GV oocytes from 59 infertile couples receiving ICSI in 2019, in Tongji Hospital, Wuhan, China. The GV oocytes derived from controlled ovarian hyperstimulation (COH) cycles underwent rescue IVM (R-IVM), and the maturation dynamics, including GVBD time (GV-MI), time from GVBD to maturation (MI-MII), maturation time (GV-MII), and MII arrest duration (MII-ICSI), were recorded by time-lapse monitoring. The matured oocytes were inseminated at different MII arrest points and subsequent embryo developments were assessed. The effects of baseline clinical characteristics, oocyte diameters, and maturation dynamics on the developmental competence of the oocytes were also analyzed. Results Totally, 157 GV oocytes were collected. GVBD happened in 111 oocytes, with a median GV-MI duration of 3.7 h. The median MI-MII duration was 15.6 h and the median GV-MII duration was 19.5 h. The maturation rate reached 56.7% at 24 h and 66.9% at 48 h, and the clinical factors, including patient age, FSH level, AMH level, ovarian stimulation protocol, and serum estradiol and progesterone levels on hCG trigger day, showed no effects on the 24-h maturation rate. The normal fertilization rate of oocytes resuming meiosis within 8 h and matured within 24 h was significantly higher than that of oocytes resuming meiosis after 8 h and matured after 24 h. Furthermore, among those oocytes matured within 24 h, the high-quality embryo formation rate of oocytes resuming meiosis within 4.5 h and matured within 19 h was significantly higher. All stated time was measured from the start point of IVM. Additionally, for oocytes from patients with serum progesterone levels less than 1 ng/ml on hCG trigger day, the high-quality embryo formation rate was significantly increased. Conclusion R-IVM technology could increase the available embryos for patients in routine COH cycles, but excessive culture beyond 24 h is not recommended. GV-MI duration of the oocyte, recorded by time-lapse system, and serum progesterone levels of patients on hCG trigger day can significantly affect the developmental potential of the IVM oocytes.

Exploration of the value of progesterone and progesterone/estradiol ratio on the hCG trigger day in predicting pregnancy outcomes of PCOS patients undergoing IVF/ICSI: a retrospective cohort study

Background Polycystic ovary syndrome (PCOS) is a common endocrine disorder with the disorders of estrogen(E2) and progesterone(P) secretion. The purpose of this study was to evaluate the association between the progesterone level or progesterone/estradiol(P/E2) ratio on human chorionic gonadotropin (hCG) trigger day and the outcome of in vitro fertilization in PCOS patients and explore the value of progesterone and P/E2 ratio for predicting the clinical pregnancy. Methods The clinical data of 1254 PCOS patients who satisfied the inclusion criteria were retrospectively analyzed, including baseline characteristics such as age, body mass index, basal sex hormone levels, et al., as well as ovarian stimulation data and clinic outcome. Results The number of follicles larger than 14 mm in diameter (P < 0.001) and retrieved oocytes (P < 0.001) was greater in the high progesterone group (progesterone ≥ 0.92 ng/mL). In the high P/E2 group(P/E2 ratio ≥ 0.3), the number of follicles larger than 14 mm in diameter (P < 0.001) and retrieved oocytes (P < 0.001), as well as the rate of high-quality embryos (P = 0.040) were significantly decreased. In ultralong GnRH agonist protocol, the implantation rate(P < 0.001), hCG positive rate (P < 0.001), clinical pregnancy rate (P < 0.001) and live birth rate (P < 0.001) were all significantly higher than long GnRH agonist protocol and GnRH antagonist protocol. The clinical pregnancy rate of high progesterone group was significantly lower than that of low progesterone group in ultralong GnRH agonist (P = 0.008). The progesterone level could be used as an indicator to predict the positive clinical pregnancy (long GnRH agonist: P = 0.001; ultralong GnRH agonist: P < 0.001) except in cycles using GnRH antagonist (P = 0.169). In the ultralong GnRH agonist, the value of progesterone level in the prediction of clinical pregnancy was significantly higher than that of the P/E2 ratio (P = 0.021). Conclusions In PCOS patients, the progesterone level is associated with clinical pregnancy rate while P/E2 ratio is not. In subgroup analysis using three different COS protocols, a significant association between progesterone level and clinical pregnancy rate can be observed in the long GnRH agonist protocol and ultralong GnRH agonist protocol. The progesterone level is significantly better than the P/E2 ratio in predicting the pregnancy outcome of PCOS patients, especially in ultralong GnRH agonist cycles.

HCG Trigger After Failed GnRH Agonist Trigger Resulted in Two Consecutive Live Births: A Case Report

Background: Failed gonadotropin-releasing hormone (GnRH) agonist trigger with no oocyte retrieved during aspiration of several follicles is a rare but recurrent situation that can be rescued by the termination of the aspiration procedure, retriggering by human chorion gonadotropin (hCG), and repeated oocyte pickup 36 h later. Failed GnRH agonist trigger is frustrating and unsatisfactory, and fertility doctors must be aware of possible hCG retriggering and retained opportunity for successful cycle outcome.Objective: In this case report, we present a woman who experienced failed GnRH agonist trigger and rescue hCG retrigger followed by two consecutive live births after frozen-thawed single blastocyst transfers.Methods: A case report.Results: Two healthy children were born in 2018 and 2020, respectively as a result of controlled ovarian stimulation for IVF, failed GnRH agonist trigger followed by hCG re-trigger, and successful retrieval of 25 oocytes.Conclusion: Retriggering with hCG after failed GnRH agonist trigger can result in consecutive live births, and such knowledge can prevent cycle cancellation and patient discouragement. Knowledge on retriggering with hCG and consecutive live births after failed GnRH agonist trigger can prevent cycle cancellation and patient discouragement.

Early Follicular Phase Human Chorionic Gonadotropin Addition May Improve the Outcomes of In Vitro Fertilization/Intracytoplasmic Sperm Injection in Patients With “Unpredictable” Poor Response to Gonadotropin-Releasing Hormone Antagonist Protocol

PurposeTo compare the effects of early and mid-late follicular phase administration of 150 IU of human chorionic gonadotropin (hCG) on gonadotropin-releasing hormone (GnRH) antagonist protocol in “unpredictable” poor ovarian response (POR) women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) treatment.MethodsA retrospective single-center cohort study was conducted on 67 patients with “unpredictable” POR in their first IVF/ICSI cycle receiving GnRH antagonist protocol. Patients were treated with a second IVF/ICSI cycle using the same GnRH antagonist protocol with the same starting dose of recombinant follicle-stimulating hormone (rFSH) as the first cycle; a daily dose of 150 IU of hCG was administrated on either stimulation day 1 (Group A, n = 35) or day 6 (Group B, n = 32). The number of oocytes retrieved, number of usable embryos, serum level of estradiol (E2) on day of hCG trigger, and clinical pregnant outcomes were studied.ResultsThe addition of 150 IU of hCG on either the first day or sixth day of stimulation increases the serum level of E2, luteinizing hormone (LH), and hCG on the day of hCG trigger. Only the use of 150 IU of hCG on the first stimulation day improved the number of oocytes retrieved, mature of oocytes, and usable embryos, but not the addition of hCG on stimulation day 6. Implantation rate, clinical pregnancy rate, and ongoing pregnancy rate showed an increasing trend in patients receiving 150 IU of hCG in the early phase compared with mid-late phase, even thought there was no statistically significant difference.ConclusionsOur study demonstrated that adding 150 IU of hCG in subsequent GnRH antagonist cycle in “unpredictable” poor responders is associated with the improvement of response to stimulation. Furthermore, early follicular phase addition of 150 IU of hCG significantly increased the number of oocytes retrieved and usable embryos than did the mid-late addition of the same dose.

Correlation between estradiol levels on day of HCG trigger and the number of mature follicles, number of oocytes retrieved, and the number of mature oocytes (M2) after oocyte aspiration in ICSI cycles

Background Estradiol is an important marker of ovarian response to ovarian stimulation in ART cycles. The study tries to find the correlation of serum estradiol on the day of HCG trigger to the number of follicles, the number of oocytes retrieved, and the number of mature oocytes, and also, to correlate estradiol per follicle and estradiol per oocyte on the day of HCG, to the number of oocytes retrieved, and to the number of mature oocytes. It is a cross sectional study using retrospective data. Results The data of 232 patients were analyzed. Our study showed a positive correlation between estradiol levels and the number of follicles (NF) (r = 0.592, p < 0.001), number of retrieved oocytes (NRO) (r = 0.576, p < 0.001), and number of mature oocytes (NMO) (r = 0.554, p < 0.001). E/follicle ratio did not have a significant correlation with NRO and NMO. E/Oocyte ratio had a strong negative correlation with NMO (r = −0.280, p < 0.001) Conclusions Serum estradiol had a positive correlation with NF, NRO, and NMO. But E/O had a strong negative correlation with NMO. These results indicate that estradiol levels can be used as an important clinical tool in the prediction of oocyte and mature oocyte yield in ART cycles. Reproductive outcome in ART cycles is largely dependent on the number of oocytes and mature oocyte yield. Estradiol levels on the day of HCG appear to strongly correlate with the outcome of ART cycles.

Predictive Value of Decreased Serum Estradiol Levels Following Human Chorionic Gonadotropin Trigger For Unexpected Ovulation During In Vitro Fertilization Cycle of Patients With Poor Ovarian Response

Background Unexpected spontaneous ovulation typically occurs in patients with poor ovarian response (POR); however, reliable predictors are lacking. We analyzed explore the predictive value of decreased serum estradiol (E2) levels on the day after human chorionic gonadotropin (hCG) trigger during in vitro fertilization (IVF) cycles. Methods We retrospectively analyzed clinical data (n = 978) in IVF cycles between 2017 and 2020. According to decreased ratios of E2 levels (△E2%) between day 1 and day 0 of hCG trigger, patients were divided into: ≤10% (group A), 10–20% (group B), and ≥ 20% (group C). Basic characteristics and laboratory data were compared using analysis of variance or Mann–Whitney U-test. Logistic regression and receiver operating characteristic (ROC) curve analyses were used to assess the relationship between △E2% and spontaneous unexpected ovulation. Fresh embryo transfer cycles in groups A, B, and C were subgrouped as A1, B1, and C1, respectively; clinical outcomes were compared. Results Mean E2 and luteinizing hormone (LH) levels, ratio of premature LH peak and high progesterone levels, time interval between hCG trigger and oocyte retrieval, number of follicles ≥ 11 mm in diameter, and dose regimen of gonadotropin among the three groups showed significant differences (P < 0.05). After adjusting for relevant confounders, likelihood ratios of unexpected ovulation in groups B and C were 2.96 (95 % confidence interval [CI]: 0.64–13.70), and 11.74 (95 % CI: 3.09–44.62), respectively, compared with that in group A. The test for trend was significant before and after correction (P for trend < 0.05). On combining premature LH peak (> 10 mIU/mL) or high progesterone level (> 1.0 ng/mL) on the day of hCG, area under ROC curve (AUC) was 0.849 (95 % CI, 0.764–0.934, P < 0.05). However, when we only considered △E2%, AUC was 0.738 (95 % CI. 0.562–0.914, P < 0.05). Rates of embryo implantation, clinical pregnancy, live birth in fresh embryo transfer cycle, and early abortion among the three subgroups did not differ significantly (P > 0.05). Conclusions E2 levels on the day after hCG trigger are valuable for predicting unexpected ovulation. However, their decreased levels do not directly affect clinical pregnancy outcomes.

Conception rate with or without hCG trigger in clomiphene induced cycles

Background: Clomiphene citrate (CC) is still the most common drug for ovulation induction. Most physicians use hCG trigger routinely for follicle rupture. Ideally hCG is recommended only where there is no spontaneous LH surge. Hence this study was conducted to see the role of hCG for follicle rupture in CC induced cycle. Aims and objectives of the study were to compare the ovulation rate in CC induced cycle with or without hCG trigger and finally the conception rate.Methods: Study was conducted in the department of OBG, LLRM medical college Meerut on women with anovulatory infertility. All women were given 50-100 mg CC. Follicular study from D-9 was done till follicle rupture. Women in group A were observed without any trigger and women in group B were given inj. hCG trigger 10,000 IU when follicle size reached 20-22 mm.Results: Conception rates were 25% Vs 31% in group A and group B. Follicle rupture was seen in 84% cases in group A and 71% in group B. Results were comparable in both the groups.Conclusions: By adding inj. hCG for ovulation trigger does not increase the conception rate. LH surge is already there in CC induced cycles. It is having role only in cases where no LH surge is there.

Metabonomics Study on the Infertility Treated With Zishen Yutai Pills Combined With In Vitro Fertilization-embryo Transfer

Zishen Yutai Pills (ZYP) is a safe and well quality-controlled TCM preparation with promising effects in many fields of reproduction, including prevention of miscarriage, increase of pregnancy rate during in vitro fertilization-embryo transfer (IVF-ET). The plasma of patients was collected from a clinical trial, namely, “Effect of Traditional Chinese Medicine vs placebo on live births among women undergoing in vitro fertilization, a multi-center randomized controlled trial.” Plasma samples were analyzed with metabonomics method. UPLC-MS technology was used to establish the plasma metabolic fingerprint. Multivariate statistical analysis was applied for comparing the differences of plasma metabolites between ZYP group and placebo group, 44 potential metabolites were screen out and identified. Pathway analysis was conducted with database mining. Compared with placebo, chemicals were found to be significantly down-regulated on HCG trigger day and 14 days after embryo transplantation, including trihexosylceramide (d18:1/26:1), glucosylceramide(d18:1/26:0), TG(22:6/15:0/22:6), TG(22:4/20:4/18:4). Compared with placebo, some chemicals were found to be significantly up-regulated on HCG trigger day and 14 days after embryo transplantation, i.e., PIP3(16:0/16:1), PIP2(18:1/18:1), tauroursodeoxycholic acid, L-asparagine, L-glutamic acid, kynurenic acid, 11-deoxycorticosterone, melatonin glucuronide, hydroxytyrosol. These metabolites were highly enriched in pathways including sphingolipid metabolism, alanine, aspartic acid and glutamic acid metabolism, aminoacyl tRNA biosynthesis, taurine and hypotaurine metabolism. This study revealed metabolic differences between subjects administered with ZYP and placebo. Relating metabolites were identified and pathways were enriched, providing basis on the exploration on the underlying mechanisms of ZYP combined with IVF-ET in the treatment of infertility.