Low-quality Embryos Research Articles

P–256 Time Lapse Imaging (TLI) acquired morphokinetic variables, nucleation errors and cleavage abnormalities are associated with live birth and may aid in de-selection of transfer embryos

Abstract Study question May the observation by TLI of morphokinetics, nucleation errors and cleavage abnormalities assist in de-selecting embryos before embryo transfer? Summary answer The combine predictive power of the association between the three biomarkers and live birth may aid in embryo de-selection What is known already Morphokinetic parameters and cleavage biomarkers are associated with treatment outcomes following in vitro fertilization (IVF). Nucleation errors observed by TLI have also been associated with IVF outcomes. It is also shown that nucleation error self-repair in pre-implantation embryos occurs, resulting in euploid blastocysts and live births. Biomarkers identified by TLI have been incorporated in developing algorithms to be used in selecting “the embryo” with the best potential for a live birth. However, the few randomized control studies (RCT) have not shown convincingly that TLI significantly improves live birth rate. Study design, size, duration Analyses of TLI data from transferred embryos, cultured in the EmbryoScope TM between June 2012 and August 2018, in a single IVF clinical setting were included. 2082 treatment cycles with Known Implantation Data (KID) for implantation and live birth were included in the analyses. Nucleation errors such as micronucleation, binucleation, and multinucleation were systematically annotated. Cleavage abnormalities such as direct cleavages, rapid and reverse cleavages were annotated for a minimum of 44 hours post insemination. Participants/materials, setting, methods Annotations for cleavage abnormalities, morphokinetic variables and nucleation errors, during a minimum of 44 hours, for 2959 transferred embryos were obtained from the EmbryoScope. The potential negative association between day 2 KID embryo biomarkers and implantation as well as live birth was assessed. The analyses controlled for potential confounding by adjusting for maternal age, infertility diagnosis, BMI, hormonal stimulation regime and insemination method. Main results and the role of chance Preliminary results were obtained regarding biomarkers in the form of nucleation errors, cleavage abnormalities and early embryo morphological attributes. Several of these biomarkers were significantly associated with implantation and live birth. Nucleation errors were associated with substantial decrease in implantation and live birth, but contrary to findings from other studies, none of the recorded nucleation error types precluded live birth. Many morphokinetically defined cleavage abnormalities were also shown to be significantly associated with implantation and live birth, with timings to 2-cells (t2) and second cell cycle (cc2) displaying the most prominent predictions for live birth probability. Within each of the three biomarker groups, logistic regression models with implantation and live birth probability predictions displayed reasonable explanatory power regarding implantation and live birth. Combining all types of biomarkers lead to logistic regression models with substantially higher explanatory power than when the regression models only comprised a single biomarker group. With a study of this size and P values for the basic findings predominantly being highly significant, the role of chance is likely to be limited. The statistical uncertainty may therefore be subordinated to the confounding caused by embryo transfer selection and further by exclusive use of embryos with known implantation Limitations, reasons for caution Only transferred embryos with KID data were analysed and hence the outcome of other embryos is unknown. Our study used mostly day 2 embryos, therefore generalisation up to blastocyst stage is not possible. Our findings apply to our study cohort and may differ from findings in another clinical setting. Wider implications of the findings: Our study provides knowledge about the role of TLI biomarkers and their potential for deselecting embryos for transfer. This will avoid transfer of lower quality embryos with lower chances of live birth. Incorporating such non-invasive de-selection strategies, alongside morphology may contribute to improving IVF outcome. Trial registration number NA

Do Seminal Isoprostanes Have a Role in Assisted Reproduction Outcome?

F2-isoprostanes (F2-IsoPs), stereoisomers of prostaglandin F2α generated by the free radical-induced oxidation of arachidonic acid, have been associated with different male infertility conditions. This study aimed to evaluate the role of seminal isoprostane levels and sperm characteristics in the reproductive outcome and embryo quality of 49 infertile couples. Semen analysis was performed following WHO guidelines. Sperm chromatin maturity was detected using an aniline blue (AB) assay, and DNA integrity was assessed using the acridine orange (AO) test. Seminal F2-IsoP levels were quantified by gas chromatography/negative ion chemical ionization tandem mass spectrometry (GC/NICI–MS/MS) analysis. Correlations among variables and their impact on in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) outcome were investigated. F2-IsoP levels are positively correlated with double-stranded DNA sperm (p < 0.001) and negatively correlated with mature sperm chromatin (p < 0.001). Patients with positive outcomes had an increased percentage of sperm with double-stranded DNA, as did patients producing high-quality embryo, who showed higher F2-IsoP levels compared to those detected in the low-quality embryo group. An intriguing relationship between a mild increase in F2-IsoP levels, DNA integrity, and embryo quality seems to indicate that the non-enzymatic oxidation of arachidonic acid can be also a marker of metabolic activity in human semen.

The Impact of Smoking on Semen Quality and ICSI Outcome in the Obese Men

Background: Male obesity with or without smoking and raised male BMI can markedly harm quality embryo blastocyst development, clinical pregnancy rate, and live birth rate in couples who underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Male BMI could be an essential factor influencing IVF–ICSI outcome. Objective: This study aimed to investigate the effect of smoking on semen quality, fertilization, embryo quality, and their impact on pregnancy after ICSI in overweight men. Material and methods: The study was performed on 150 cases in the assisted reproductive unit in International Islamic Center for Population Studies and Research, Al-Azhar University. The recruited males were divided into three groups (50/group): Normal control (C), Overweight (OW), and overweight with a smoking (OW&S). Their physiological, biochemical, and semen parameters were investigated for all cases. The fertilization rate, cleavage rate, embryo quality, and pregnancy rate were examined and recorded during and after the ICSI process. Results: Percent of abnormal sperm morphology in the OW&S group showed a very highly significant increase (P<0.01) than the control group. However, for the overweight with smoking (OW&S) and overweight groups (OW), the percentage of the fertilization rate and the high-quality embryos were significantly lower than the values of the control group (P<0.01). The percentage of the low-quality embryo was higher in OW&S and OW groups than in the control group. The lowest rate of positive pregnancy test recorded in overweight with smoking (46%) followed by overweight (60%). Conclusion: the semen parameters and hormonal profile, fertilization rate, embryo quality, and pregnancy rate were significantly lower in smoker men suffering from overweight.

THE USE OF THE PRP THERAPY TO ACTIVATE FOLLICULOGENESIS IN WOMEN OF THE LATE REPRODUCTIVE AGE IN PROGRAMS OF ASSISTED REPRODUCTIVE TECHNOLOGIES

The rate of infertility in married couples of reproductive age in this country makes up from 10 to 15%, in some regions this value is close to 20% - acritical level that has a negative impact on demographic figures. The rate of pregnancy depends directly on the women’s age and decreases by 3.3times starting from the age of 19 and by the age of 48. The decrease in the ability to conceive is accounted for by subtle mechanisms related to thedeterioration of the quality of oocytes. The patients who are prepared for an extracorporeal fertilization program (ECF) and do not respond tocontrolled ovarian hyperstimulation protocols are considered to be poor respondents. Many studies focused on the development of an optimaltreatment method. However, none of the approaches seems to be effective enough to guarantee a successful use. Platelet-rich plasma is a new andpromising method that is successfully used in the reproductive science to solve a number of medical problems. All the patients whose commonfeatures were low oocyte output and poor embryo quality as well as failed ECF attempts were offered a treatment method of autologous PRP therapyafter their written consent. Three months later, one obtained astonishing results, which by the markers of biochemical infertility alone could beclassified as a complete biological phenomenon and are also characterized by improved embryo quality. The results of hormonal homeostasis show adecrease in the level of the follicle-stimulating hormone by 67.33% while the level of the anti-muellerian hormone is 75.18% higher.Thus, the use of the PRP therapy in poor respondents helps them to overcome their problematic reproductive barrier.

Evaluation of the In Vitro Fertilization Success Rate in Transfer of Top-Quality Embryo Versus Poor-Quality Embryos: A Cohort Study

Objectives: To evaluate the in vitro fertilization success rate by transferring top- versus low-quality embryos. Materials and Methods: This prospective cohort study was conducted on 199 infertile women. Ninety-nine patients underwent an agonist cycle (70 fresh embryo transfer [ET] and 29 frozen ET), and 100 patients received an antagonist cycle (28 fresh ET and 72 frozen ET) in the infertility department of a tertiary university-based hospital between May 2019 and March 2020. The blastocysts classified as AA, AB, and BB, as well as AC, BC, and CC were considered as top- and poor-quality embryos (TQE and PQE). The study outcomes were biochemical and clinical and determined the rate of pregnancy. Results: The average age of the participants was 32.44±5.25 years old. Women with TQE were significantly younger than those with PQE (31.35±4.97 vs. 34.09±5.27, P&lt;0.001). In addition, the duration of women’s infertility was significantly (P&lt;0.001) correlated with the embryo’s top quality. A positive β-human chorionic gonadotropin was detected in 12.6% (n=25) of women while clinical pregnancy was investigated in 8% (n=16) of them. The fetal heart rate was detected in 7.5% (n=15). Eventually, the clinical (P=0.020) and determined (P=0.030) pregnancy rates significantly differed between two study groups with a higher level in the TQE group. Conclusions: It seems that TQP transfer should be the first recommendation for infertile women, but when the double-embryo transfer (DET) is needed according to the patient’s condition, she should be informed that the quality of the second embryo may have an adverse impact on pregnancy consequences.

Embryo biosensing by uterine natural killer cells determines endometrial fate decisions at implantation.

Decidualizing endometrial stromal cells (EnSC) critically determine the maternal response to an implanting conceptus, triggering either menstruation‐like disposal of low‐fitness embryos or creating an environment that promotes further development. However, the mechanism that couples maternal recognition of low‐quality embryos to tissue breakdown remains poorly understood. Recently, we demonstrated that successful transition of the cycling endometrium to a pregnancy state requires selective elimination of pro‐inflammatory senescent decidual cells by activated uterine natural killer (uNK) cells. Here we report that uNK cells express CD44, the canonical hyaluronan (HA) receptor, and demonstrate that high molecular weight HA (HMWHA) inhibits uNK cell‐mediated killing of senescent decidual cells. In contrast, low molecular weight HA (LMWHA) did not attenuate uNK cell activity in co‐culture experiments. Killing of senescent decidual cells by uNK cells was also inhibited upon exposure to medium conditioned by IVF embryos that failed to implant, but not successful embryos. Embryo‐mediated inhibition of uNK cell activity was reversed by recombinant hyaluronidase 2 (HYAL2), which hydrolyses HMWHA. We further report a correlation between the levels of HYAL2 secretion by human blastocysts, morphological scores, and implantation potential. Taken together, the data suggest a pivotal role for uNK cells in embryo biosensing and endometrial fate decisions at implantation.

The Impact of Down-Regulation on Obstetrics and Perinatal Outcomes in Singleton Pregnancies After In Vitro Fertilization.

Backgrounddown-regulation has been widely used in IVF treatment; however, it lacks reports on the impact of down-regulation on obstetrics and perinatal outcomes. The purpose of this study was to compare the obstetrics and perinatal outcomes among different down-regulation conditions.Methodsthis is a retrospective cohort study on 3578 patients achieving cumulative singleton clinical pregnancy after their first oocytes retrieval cycle. Patients were grouped according to the serum estradiol after down-regulation (E2D) into three groups: <30, 30-55, >55 pg/ml. The obstetrics and perinatal outcomes, and live-birth rate per clinical pregnancy were main outcome measures. In the subgroup analysis, patients were further divided according to the mode of transfer. ANOVA, chi-square test, multivariate logistic regression, and multivariate general linear model were performed for statistical analysis.Resultsthe patients with E2D <30, 30-55, >55 pg/ml had similar live-birth rates. The patients with E2D <30 pg/ml had a lower risk of hypertension disorders than those with E2D 30-55 pg/ml. No difference was found in the risks of placenta previa, placenta abruption, premature rupture of membrane, hemorrhage, gestational diabetes mellitus, or intrauterine growth restriction. The newborns in the group with E2D <30 pg/ml had a lower risk of PICU admission than those in the group with E2D >55 pg/ml. There was no difference in the risks of congenital anomalies or mortality among the three groups. No differences were found in the gestational week, percentages of preterm birth and very preterm birth, birth weight, percentages of low birth weight and very low birth weight, delivery mode, or sex of newborn. Subgroup analysis showed that E2D 30-55 pg/ml was associated with a higher risk of low birth weight in patients with one fresh transfer + frozen transfer(s).ConclusionDown-regulation has no effect on the live-birth rate per clinical pregnancy. Patients with E2D <30 pg/ml may have advantages regarding lower risks of both maternal hypertension and newborn PICU admission. E2D 30-55 pg/ml may be associated with low birth weight in patients with relatively low quality embryos.

Adding a low-quality blastocyst to a high-quality blastocyst for a double embryo transfer does not decrease pregnancy and live birth rate.

The effect of embryo quality on clinical outcomes of assisted reproductive technology following a double transfer is not well defined, with some studies suggesting that a low-quality embryo transferred with a high-quality embryo decreases the live birth rate (LBR), compared with transferring a single high-quality embryo. Our study examined whether the quality of a second blastocyst transferred affects the outcome, controlling for the number of the available high-quality blastocysts (HQB). A historical cohort study of 2346 fresh blastocyst transfers in a single fertility clinic between 2013 and 2019. The main outcomes were pregnancy, miscarriage, live birth, and multiple gestation rates. Outcomes were compared between single embryo transfers with a high-quality blastocyst (SET-H), double embryo transfers with two HQBs (DET-HH), and transfers with one high-quality and one low-quality blastocyst (DET-HL). Outcomes were also assessed between SET and DET when only low-quality blastocysts were available. With one HQB available, DET-HL increased LBR (adjusted odds ratio [OR] 1.65, 95% CI 1.09-2.49) compared with SET-H, but increased multiple gestation rate (aOR 23.1, 95% CI 3.0-177.6). With two HQBs available, DET-HH was associated with a higher LBR (aOR 1.62, 95% CI 1.28-2.04) and lower miscarriage rate (aOR 0.56, 95% CI 0.40-0.80), but very high twin rate (aOR 49.8, 95% CI 24.3-102.1) compared with SET-H. A SET-H with at least one or more HQB available to freeze, compared with a SET-H with no other HQB available, had a higher LBR (aOR 1.69, 95% CI 1.17-2.45). When there were no HQBs available, compared with SET-L, a DET-LL had a higher live birth (aOR 3.20, 95% CI 1.78-7.703) and twin rate (aOR 3.72×1010 ) and a lower miscarriage rate (aOR 0.24, 95% CI 0.10-0.58). When there is one HQB available, transferring an additional low-quality blastocyst would only slightly increase the LBR, but significantly increase the twin rate, therefore SET should be recommended. When two or more HQBs are available, SET-H would have a reasonably good chance of success without the very high twin rate associated with DET-HH. DET-LL when compared with SET-L, would increase the LBR, but increase the risk of multiple gestation.

Effect of Embryo Developmental Stage, Morphological Grading, and Ploidy Status on Live Birth Rate in Frozen Cycles of Single Blastocyst Transfer.

To determine whether embryo developmental stage or morphological grading can predict live birth rate (LBR) from a single blastocyst in nonbiopsied and biopsied frozen embryo transfer (FET) cycles. This retrospective study included 1336 nonbiopsied and 360 euploid FET cycles. Blastocysts were divided according to developmental stage (day 5 [D5] and day 6 [D6]) and morphology (good quality and low quality). Nonbiopsied cycles in which D5 blastocysts were transferred were associated with a significantly higher LBR than those in the D6 group (48.5 vs. 24.3%; p< 0.001), as well as in good-quality embryo transfer cycles than that in low-quality embryo cycles (52.6 vs. 25.3%; p< 0.001). Embryos reaching good-quality blastocysts on D5 yielded significantly higher LBR than those similar quality blastocysts on D6. The same trend was seen in low-quality embryos. Concerning only D5 or D6 blastocyst transfer, the LBRs of good-quality embryos were still superior to those of low-quality embryos. In the case of euploid embryo transfers, the LBR (48.9 vs. 44.9%, p= 0.444) of D5 blastocysts did not significantly differ from that of D6 blastocysts. Good-quality embryos showed a higher LBR than low-quality embryos (51.6 vs. 40.0%, p= 0.030); the adjustedodds ratio remained insignificant after controlling for confounders (aOR 1.56; 95% CI 0.99-2.45; p= 0.056). The LBRs in the same developmental stage or morphology subgroups were not statistically significant. Embryo developmental stage and morphological grade are useful predictors of LBR in nonbiopsied FET cycles. However, no association was found in euploid transfer cycles.

ASSOCIATION BETWEEN STANDARD MORPHOLOGY ASSESSMENT AND CHROMOSOME CONSTITUTION AFTER NGS ANALYSIS

To determine the correlation between standard morphology assessment and chromosome constitution in blastocyst analyzed by NGS. Retrospective observational study Data was collected retrospectively from June 2016 to October 2019. In 1221 IVF cycles, 4719 blastocysts were biopsied and analyzed by NGS. DNA from trophectoderm cells were amplified by using Sureplex, hr-NGS was performed with VeriSeq PGS assay, (MiSeq-Ilumina). NGS data was analyzed with Bluefuse sofware (Ilumina). Chromosome constitution was classified according to the genetic results as: euploid, single aneuploid, double aneuploid, complex aneuploidy (>3 chromosomes are aneuploid), (single, double and complex) mosaic, complex mix and mix (aneuploid and mosaic). Embryo morphology was classified as good, fair, or poor based on SART. Descriptive statistics and multilevel model for logistic regression considering a patient as second level and blastocyst as first level were used to evaluate the association between chromosome constitution and embryo morphology which was adjusted by levels of trophectoderm and inner cell mass, maternal age, fertilization technique and day of embryo biopsy. p<0.05 as significant level. All analyses were performed by Stata. For all blastocyst analyzed the frequency was euploid (43.6%), single aneuploid (14.2%), double aneuploid (4.7%), complex aneuploid (2.4%), single mosaic (14%), double mosaic (4.9%), complex mosaic (7.3%), mix (2.9%) and complex mix (5.3%). Most of the embryos biopsied have a good (46.8%) and fair (47.9%) quality and 5.3% were poor quality. Euploidy embryos were associated to have good morphology as is shown in table 1. Interestingly, poor embryo quality have a significantly increased frequency of double aneuploidy (p=0.013). On the contrary, any associations were found for double mosaic, complex aneuploid and mix.Tabled 1Table 1. Association between chromosome constitution and embryo morphologyGoodFairPoorEuploid1092 (53.06%)880 (42.76%)86 (4.18%)OR10.74 (0.65-0.84)**0.70 (0.52-0.94)*Aneuploid301 (44.93%)331 (49.40%)38 (5.67%)OR10.99 (0.82-1.19)0.94 (0.64-1.39)Mosaic290 (43.81%)339 (51.21%)33 (4.98%)OR11.25 (1.03-1.51)*1.25 (0.82-1.89)Double Aneuploid86 (39.09%)109 (49.55%)25 (11.36%)OR11.04 (0.75-1.44)1.94 (1.15-3.26)*Complex Mix79 (31.60%)147 (58.80%)24 (9.60%)OR11.54 (1.13-2.10)**2.03 (1.24-3.33)**OR (IC 95%): Odds Ratio (Confident Interval 95%),* p<0.05, ** p<0.001. Open table in a new tab OR (IC 95%): Odds Ratio (Confident Interval 95%),* p<0.05, ** p<0.001. NGS results confirmed the association between embryonic morphology and euploidy. Mosaicism was reported more frequently in fair embryos, while double aneuploidy and mixed complex are more associated with embryos with low embryo quality.

Pretreatment with dienogest in women with endometriosis undergoing IVF after a previous failed cycle

Research questionThe major causes of IVF failure in women with endometriosis have been attributed to decreased ovarian reserve, low embryo quality and impaired receptivity of the endometrium. Dienogest (DNG) has anti-inflammatory and anti-angiogenic activity and so may theoretically improve IVF outcomes in women with endometriosis. This study aimed to evaluate the administration of DNG before IVF in women with endometriosis who had previously failed one IVF cycle. MethodsThis study was based on the retrospective analysis of a prospectively collected database, including 151 women who had failed a previous IVF cycle and all subsequent embryo transfers and had an imaging diagnosis of endometriosis. Patients either directly underwent IVF without receiving hormonal treatment or received 3 months of treatment with DNG (2 mg/daily) before IVF. ResultsEighty-eight (58.3%) patients underwent IVF without previous hormonal treatment, and 63 (41.7%) received pretreatment with DNG. The cumulative implantation, clinical pregnancy and live birth rates were significantly higher in the DNG-treated group (39.7%, 33.3% and 28.6%) than in the non-treated group (23.9%, 18.2% and 14.8%; P = 0.049, 0.037 and 0.043, respectively). The largest diameter of endometriomas significantly decreased after DNG pretreatment (P < 0.001). The use of DNG increased significantly the number of oocytes retrieved (P = 0.031), two-pronuclear embryos (P = 0.039) and blastocysts (P = 0.005) in women with endometriomas of diameter ≥4 cm. ConclusionsThis study suggest that in patients with endometriosis, IVF outcomes can be improved by pretreatment with DNG. In particular, the use of DNG allows for better oocyte retrieval and blastocysts in patients with large endometriomas.

Is transferring a lower-quality embryo with a good-quality blastocyst detrimental to the likelihood of live birth?

To determine if transferring a lower-quality embryo with a good-quality blastocyst is detrimental, given that evidence suggests that embryos can signal the endometrium and that embryo quality may affect negatively endometrial receptivity. Retrospective cohort study. Invitro fertilization center. Single- versus double-embryo transfer. Patients with a double-embryo transfer of a good-quality blastocyst plus a lower-quality blastocyst, early blastocyst, or morula were compared with patients receiving a single good-quality blastocyst. Live birth, multiple gestation. In this study, 4,640 invitro fertilization cycles were analyzed. In none of the analyses did transferring a second lower-quality embryo negatively affect birth rate. In the primary analysis, transferring a second lower-quality embryo increased live birth by 10% and the multiple birth rate by 15%. The addition of a fair- or poor-quality blastocyst or early blastocyst markedly increased the twin birth rate by 22%-27% with an 8%-12% increase in live birth. The addition of a morula did not increase live birth but resulted in 12% more multiples. In women younger than 38 years, adding a lower-quality embryo increased the birth rate by 7% but resulted in 18% increase in multiples. In women 38 years or older, adding a lower-quality embryo increased the live birth rate by 9% with a 15% increase in multiples. Addition of a lower-quality embryo does not have a detrimental effect on a good-quality blastocyst and results in a small increase in live births. However, this is at the expense of a marked increase in the likelihood of multiple gestations.

PERFIL DOS CASAIS INFÉRTEIS SUBMETIDOS À TÉCNICA DE REPRODUÇÃO HUMANA ASSISTIDA

&#x0D; O objetivo deste estudo foi caracterizar o perfil dos casais inférteis submetidos à técnica de reprodução humana assistida (RHA) no município de Salvador, Bahia. Trata-se de um estudo de caráter retrospectivo, observacional, baseado em dados secundários, obtidos mediante consulta aos prontuários eletrônicos dos pacientes cadastrados em uma clínica particular de RHA durante o ano de 2017. As variáveis categóricas e quantitativas foram apresentadas em valor absoluto e relativo, média ± desvio-padrão, respectivamente. Posteriormente, analisadas por meio de um software estatístico. Dos 496 pacientes atendidos na clínica de reprodução, 360 (72,6%) foram diagnosticados com infertilidade. A causa compartilhada (n=224; 62,2%) foi mais prevalente do que a isolada por sexo. Entre as mulheres, observa-se que a idade avançada (30,8%) e endometriose (26,5%) representam as principais causas de infertilidade. Nos homens, a teratozoospermia (36,9%) e a varicocele (33,6%) foram os fatores que mais comprometeram a fertilidade. A qualidade do gameta feminino e masculino é severamente comprometida em pacientes que tem como causa de infertilidade a falência ovariana (62,5%) e oligoastenoteratozoospermia (56,5%), respectivamente. No entanto, observa-se que nas mulheres, a baixa qualidade embrionária está mais associada à obstrução tubária (47,7%), enquanto que nos homens, o mesmo fator associado a baixa qualidade seminal é também responsável pelo desenvolvimento insatisfatório dos embriões (69,2%). Por fim, conclui-se que a infertilidade acomete, em maior proporção, homens e mulheres simultaneamente. Nas mulheres, a infertilidade associa-se a idade avançada e endometriose, enquanto que nos homens a teratozoospermia e varicocele. Essas causas, além de impactarem na qualidade dos gametas, também afetam a qualidade e desenvolvimento embrionário.&#x0D; &#x0D;

Lipid droplets in granulosa cells are correlated with reduced pregnancy rates

BackgroundLipids are an important source for energy production during oocyte maturation. The accumulation of intracellular lipids binds to proteins to form lipid droplets. This may lead to cellular lipotoxicity. The impact of lipotoxicity on cumulus and granulosa cells has been reported. This pilot study evaluated their correlation to oocyte and embryo quality.DesignProspective case-control study.Setting: Referral IVF unit.Patients: Women younger than age 40, undergoing IVF with intracytoplasmic sperm injection.Interventions: 15 women with BMI > 30 (high BMI) and 26 women with BMI < 25 (low BMI) were enrolled. IVF outcomes were compared between groups based on BMI. Lipid content in cumulus and granulosa cells was evaluated using quantitative and descriptive methods. Lipid profile, hormonal profile and C-reactive protein were evaluated in blood and follicular fluid samples. Demographic and treatment data, as well as pregnancy rates were collected from electronic medical records.ResultsHigher levels of LDL and CRP, slower cell division rate and lower embryo quality were found in the group with high BMI. There was no difference in pregnancy rates between groups. In light of these findings, treatment outcomes were reanalyzed according to patients who became pregnant and those who did not. We found that patients who conceived had significantly lower fat content in the granulosa cells, reflected by mean fluorescence intensity recorded by flow cytometry analysis (23,404 vs. 9370, P = 0.03).ConclusionsBMI has no effect on lipid content in cumulus and granulosa cells, and does not affect likelihood of pregnancy. However, women who achieved pregnancy, regardless of their BMI, had lower lipid levels in their granulosa cells. This finding is important and further study is needed to evaluate lipid content in granulosa cells as a potential predictor of IVF treatment success.

Embryonic development kinetics after autologous bone marrow mesenchymal stem cell- derived mitochondria transfer into compromised oocytes：a prospective self-controlled study

We aimed to explore whether autologous bone marrow mesenchymal stem cell(BMSC)-derived mitochondria transfer into compromised oocytes change their embryonic development kinetics and improve outcomes in women with multiple in vitro fertilization (IVF)/ Intracytoplasmic sperm injection (ICSI) failures due to low oocyte quality A prospective self-controlled study This prospective self-controlled study was conducted at the Department of Assisted Reproduction of the sixth affiliated hospital of Sun-Yet san university from January 2018 to June 2018. Patients were voluntarily enrolled meeting the following criteria: [1] undergoing ICSI program, [2]<42 years of age, [3] body mass index <30 kg/m2, [4]at least one previous failed IVF with Low embryo quality. Low embryo quality was understood as 1) no fertilized MIIs; 2) deficient-quality embryos according to morphologic criteria ; 3) arrested embryos. Patients with abnormal chromosome were excluded. BMSCs were isolated from 20ml bone marrow and cultured. Three days before oocyte retrieval, mitochondria were isolated by differential centrifugation from BMSCs. The retrieved oocytes were randomly and averagely divided into two groups i.e. Mitochondria transfer (MT) groups and control group. In MT group, 4000-5000 copies mitochondria DNA were injected into each oocyte during intracytoplasmic sperm injection. By means of a time-lapse system (EmbryoScope; Unisense FertiliTech, Aarhus, Denmark), this study determined the timing of a number of developmental parameters including cleavage timing from a zygote to a 8-cell embryo (t2, t3, t4, t5, t6, t7, t8) and assessed fragmentation at each stage. The intervals between two consecutive cleavages were also analyzed. Duration of the second cell cycle (cc2 =t 3 – t2) is the time from the division into a two-blastomere embryo until the time to the division into a three-blastomere embryo, and second synchrony (s2 =t 4 – t3) is the time from this division into a four-blastomere embryo. A total of 25 patients were included and we got 231 oocytes in total. Their average age was 33.00 years old. The average antimullerian hormone level and antral follicles was 3.86 ng/ml and 14.68 respectively. Most of patients was primary infertility(72.22%) and the major cause of infertility was tubar factor(64.00%). We observed that the timings of all embryo cleavage stages (from t2to t8) together with fragmentation values showed no significant differences between embryos deriving from oocytes with MT or without MT. It was noteworthy that s2 was shorter in MT group, although difference didn’t reach statistical significance ( 4.20 vs. 5.90). in addition, Oocytes of MT groups had little higher fertilization rate (89.06% vs.88.35%, P= 0.865). This study demonstrated that BMSC-derived autologous mitochondria transfer didn’t alter embryonic development kinetics. It might help to improve embryo development synchrony and fertilization.

Clinical pregnancy outcome according to age of patients with high proportion of failed oocyte maturation in ICSI cases

The purpose of this study was to compare the clinical pregnancy outcomes according to age of patients with high proportion of maturation arrest oocytes in ICSI cases. Retrospective cohort study. This study was conducted on patients with ICSI cases who had transferred embryos. Clinical outcomes were analyzed by dividing the maturation arrest rate (under 40%/over 40%) and the age of 38yrs (under 38yrs/over 38yrs). From June 2011 to December 2018, a total 2495 cycles were analyzed in this study. All the patients underwent ICSI cycle followed by fresh embryo transfer. Inclusion criteria: female age between 23 and 48yrs, use of fresh or cryopreserved sperm. Exclusion criteria: surgically retrieved sperm. These patients were divided into group A (maturation arrest rate <40%, Age <38yrs), group B (maturation arrest rate ≥40%, Age <38yrs), group C (maturation arrest rate <40%, Age ≥38yrs) and group D (maturation arrest rate ≥40%, Age ≥38yrs). The pregnancy outcomes were compared among these 4 groups. A total of 2495 cycles were included (group A (n) = 1355, group B (n) = 90, group C (n) =960 and group D (n) = 54). There was no significant differences in fertilization rate between groups (group A vs. B: 80.7% ± 20.0 vs 81.0% ± 24.1, P=0.958 and group C vs. D: 83.2% ± 20.1 vs. 88.5% ± 19.6, P=0.073). But there was a significant difference in embryo quality, pregnancy rate, and miscarriage rate. More than 40% of oocyte maturation failure group had lower levels of embryo quality. There was significant difference in at least one good quality embryo transfer cycle rate (group A vs. B: 69.4% vs. 46.7%, P<0.001 and group C vs. D: 63.1% vs. 55.6%, P<0.001). And more than 40% of oocyte maturation failure group had lower clinical pregnancy rate (group A vs. B: 41.6% vs. 20.0%, P<0.001 and group C vs. D: 25.7% vs. 7.4%, P<0.001) and ongoing pregnancy rate (group A vs. B: 35.9% vs. 14.4%, P<0.001 and group C vs. D: 17.0% vs. 1.9%, P<0.001). According to our study, high rate of oocyte maturation failure group had lower embryo quality, pregnancy rate and higher miscarriage rate. However, there was no effect on in fertilization rate. There was a similar tendency in analysis according to age. High clinical results can be maintained only by lowering the proportion of mature failed oocyte as much as possible.

Cell-free nucleic acids and melatonin levels in human follicular fluid predict embryo quality in patients undergoing in-vitro fertilization treatment

Despite many advances in assisted reproductive technology (ART), the most viable embryo selection remains a challenge for infertility treatment. This study was designed to investigate whether intra-follicular circulating cell-free DNA (cfDNA) fragments and Melatonin levels predict embryo quality in patients undergoing IVF treatment. A total of eight hundred and ninety-five follicular fluid (ff) samples were collected from 325 infertile patients undergoing IVF treatment. Patients were enrolled from August 2017 to December 2018 in the infertility center of a tertiary care hospital. A clear non-hematic follicular fluid was aspirated after the removal of eggs from the dominant follicles (>18mm) of each patient. Melatonin and E2 levels in each follicular sample were estimated by immune-chemiluminescence using commercially available kits. ALU-qPCR evaluated cfDNA levels in individual follicular fluid samples. Our study presented a significant and negative relationship between intra-follicular cfDNA and melatonin concentration (-0.541: P<0.001). Each individual follicle contains measurable copy number of cfDNA [mean: 1.85±2.98ng/μl (median; 1.86ng/μl (95% Cl: 0.96–2.87)]. In pregnant women cfDNA copy number was significantly decreased in follicular fluid samples(ff) aspirated from matured oocytes than in immature ones [p<0.01; β = -0.42±0.49; median; 1.45ng/ml (95% Cl: 0.36–2.97) vs. 3.57ng/μl (95% Cl: 0.37–4.01) respectively. While melatonin concentration in ff samples corresponding to mature oocytes was significantly higher than in ff samples related to immature oocytes (p<0.001). Moreover, in pregnant women cfDNA level was significantly lower in ff samples related to oocytes which produces top-quality embryos versus low quality embryos [p<0.001; β=1.81±0.91; median; 1.25ng/μl (95% Cl: 0.35–1.97)] vs. [(median; 3.65ng/ml (95% Cl: 1.23–6.36)] respectively. Likewise, in non-pregnant women melatonin levels were significantly decreased in ff samples related to embryos with high fragmentation rate (≥25%) than embryos with low fragmentation rate (<25%; p<0.001). Conclusively, this study indicates that Intra-follicular cfDNA and melatonin concentration possibly a new supplemental tool that supports to establish an advanced non-invasive early prognostic test for the patients undergoing IVF/ICSI procedure.

Impact of Oocyte Morphology on Fertilization, Embryo Quality and Pregnancy after ICSI: Evaluation with Diabetes and PCOS

Abstract Intra-cytoplasmic sperm injection (ICSI) has become an essential procedure in assisted reproductive units for infertile patients. Polycystic ovary syndrome (PCOS) and diabetes are two syndromes with their drawbacks on fertility. Objectives: The aim of this research was to study the impact of the morphological abnormalities of MII oocytes on fertilization rate, embryo quality and pregnancy following ICSI in patients with diabetes, PCOS patients. Patients and methods: This prospective observational study was done on 177 women. MII oocytes with normal or abnormal morphology, fertilization rate, embryo quality, and positive or negative pregnancies were assessed in all patients and compared between the different groups. Results: In the control group the total number of oocyte abnormalities was 146/446 MII oocytes with a percentage of (33%). In the diabetes group the total number of abnormalities in MII oocytes was 231/90 MII oocytes with a percentage of (257%), and in the PCOS the total number of abnormalities was 414/229 abnormalities in the MII oocytes with a percentage of (181%). In diabetic patients despite the higher oocytes abnormalities and the more low quality embryos than in PCOS patients, the pregnancy rate were significantly higher in the diabetes group (59.5%) compared to that in the PCOS group (51.9%),with the highest percentage of pregnancy presented in the control group (83.7%). Conclusion: ICSI outcome was significantly better in diabetic patients when compared to PCOS patients. The best outcome was observed in the control group.

Prostaglandin F2\u03b1 (PGF2\u03b1) production possibility and its receptors expression in the early- and late-cleaved preimplantation bovine embryos

BackgroundProstaglandin F2α (PGF2α) is an important component for the physiology of female reproductive processes. In the literature, the data pertaining to the synthesis and action of PGF2α in early embryonic bovine development are limited. In our study, we used the bovine in vitro culture model based on the time of first cleavage to determine the mRNA expression and immunolocalization of PGF2α synthase and its receptor in bovine embryos from the 2-cell stage to the hatched blastocyst stage. We also evaluated PGF2α production at 2, 5 and 7 days of in vitro culture.ResultsWe found a significantly higher proportion of blastocysts obtained from the early-cleaved embryos than from the late-cleaved embryos (37.7% vs. 26.1% respectively, P < 0.05). The PGFS mRNA expression was significantly higher in the late-cleaved group than in the early-cleaved group at the 2-, 4- and 16-cell stages (P < 0.05). For PTGFR, we observed that within the late-cleaved group, the mRNA abundance was significantly higher in embryos at the 2- and 16-cell stages than in embryos at the 4- and 8-cell stages (P < 0.05). We observed that PTGFR mRNA expression was significantly higher in the 2- and 16-cell embryos in the late-cleaved group than that in the early-cleaved group embryos (P < 0.05). Among the blastocysts, the PGFS and PTGFR expression levels showed a trend towards higher mRNA expression in the late-cleaved group than in the early-cleaved group. Analysis of PGF2α production showed that within the early-cleaved group, the content of PGF2α in the in vitro culture medium was significantly higher on day 7 than it was on day 2 (P < 0.05).ConclusionsThe mRNA expression levels of PGF2α synthase and its receptor depend on the developmental stage and the embryo quality. Analyses of PGFS and PTGFR expression in bovine blastocysts and of PGF2α embryo production suggest that prostaglandin F2α can act in an autocrine and paracrine manner in bovine in vitro-produced preimplantation embryos. Moreover, the tendency of PTGFR and PGFS mRNA expression to be upregulated in embryos with low developmental potential can indicate a compensation mechanism related to high PGFS and PTGFR mRNA expression levels in low-quality embryos.

A prospective study of C-reactive protein in patients with obesity during IVF

This prospective cohort pilot study investigated the physiology of C-reactive protein (CRP) during in vitro fertilization (IVF) cycles and its effect on outcomes in women with and without obesity. The study was conducted from April to August 2014, in the IVF Unit of a university-affiliated hospital. Women aged 18–42 years were enrolled. Those with chronic inflammatory diseases or acute illness were excluded. A total of 31 patients were included: 17 with BMI < 30 kg/m2 and 14 with BMI ≥ 30 kg/m2. Serum CRP levels were measured: (i) before starting ovarian stimulation; (ii) on the day of β-HCG administration; and (iii) on day of ovum pick-up (OPU), in both serum and follicular fluid. Serum CRP levels were significantly higher in women with obesity at all time points. For the entire cohort, a positive correlation was found between basal oestradiol (E2) and basal CRP (r = 0.71, p < 0.05). A specific pattern of CRP levels was not detected during the IVF cycle. High serum CRP levels on OPU day had a negative effect on embryo quality (p = 0.056). CRP ≥ 0.5 mg/dL was associated with lower quality embryos (2.6 ± 0.3 vs. 3.3 ± 0.3; p = 0.04). High serum CRP level on OPU day negatively affects embryo quality.