Embryo Score Research Articles

Simplified static embryo score system for the prediction of blastocyst formation and euploidy

Simplified static embryo score system for the prediction of blastocyst formation and euploidy

A minimally invasive methodology based on morphometric parameters for day 2 embryo quality assessment

The risk of multiple pregnancy to maternal–fetal health can be minimized by reducing the number of embryos transferred. New tools for selecting embryos with the highest implantation potential should be developed. The aim of this study was to evaluate the ability of morphological and morphometric variables to predict implantation by analysing images of embryos. This was a retrospective study of 135 embryo photographs from 112 IVF–ICSI cycles carried out between January and March 2011. The embryos were photographed immediately before transfer using Cronus 3 software. Their images were analysed using the public program ImageJ. Significant effects (P < 0.05), and higher discriminant power to predict implantation were observed for the morphometric embryo variables compared with morphological ones. The features for successfully implanted embryos were as follows: four cells on day 2 of development; all blastomeres with circular shape (roundness factor greater than 0.9), an average zona pellucida thickness of 13 µm and an average of 17695.1 µm2 for the embryo area. Embryo size, which is described by its area and the average roundness factor for each cell, provides two objective variables to consider when predicting implantation. This approach should be further investigated for its potential ability to improve embryo scoring.

Sperm DNA integrity testing: big halo is a good predictor of embryo quality and pregnancy after conventional IVF.

Sperm DNA integrity is a sperm functional parameter of male fertility evaluation. Two parameters of sperm DNA integrity were observed: DNA damage expressed as DNA fragmentation index (DFI) and percentage of the DNA undamaged spermatozoa expressed as big halo. Halosperm test was used for sperm DNA integrity determination. The aim of this study was to evaluate which DNA integrity parameter is better as an embryo quality and pregnancy prognostic parameter after the conventional IVF. We evaluated two embryo groups (positive and negative group) according to the 3rd day cumulative embryo score. Big halo and DFI, as we expected, showed good correlation (r = -0.69; p < 0.001). Receiver operating characteristic (ROC) analyses show that DFI and big halo are significant (p < 0.001) as prognostic parameters of embryo quality. ROC curves comparison of DFI and big halo revealed the AUC value for big halo to be significantly higher (DFI AUC = 0.71 vs. big halo AUC = 0.83; p = 0.025) than for DFI. Big halo was found to be the only independent predictor of embryo quality. Sperm DNA integrity both parameters are good prognostic parameters of embryo quality after the conventional IVF where big halo seems to be better. ROC analyses show DFI and big halo as significant prognostic parameters for achieved pregnancy (AUC ± SE for DFI was 0.67 ± 0.06 and 0.75 ± 0.06 for big halo). To our knowledge, this is the first study demonstrating the correlation between sperm DNA undamaged rate expressed as big halo parameter and semen characteristics as well as the influence on fertilization rate, embryo quality and pregnancy in conventional IVF.

Time-lapse embryo imaging technology

The purpose of the review is to summarize recent developments in time-lapse technologies and early embryo morphokinetics and to discuss their impact on current clinical outcomes. Contemporary embryo culture and selection methodologies that are based on classical morphology are clearly limited in providing the most suitable embryo for a successful pregnancy. Noninvasive observation of embryo development by capturing the images with a time-lapse device has recently been proposed to be a better method of embryo viability assessment. Such methodologies have been shown to increase the quality and the quantity of information on the viability without disturbing the culture conditions. Commercial availability of different time-lapse devices for human embryos facilitated the use of morphokinetics as an additional tool in human embryo selection. The application of such technologies has already shown positive results on clinical outcome by increasing our scope of traditional embryo selection, leading to higher implantation and clinical pregnancy rates. Additional benefit can come from the less-disturbed incubation environment that is created by all-in-one incubators. Such devices can also be very important research tools in order to observe and analyze the effect of different patient-specific or clinical conditions on embryo development parameters that are not available through classical embryo scoring.

The association between embryo quality and perinatal outcome of singletons born after single embryo transfers: a pilot study.

Does the quality of a single transferred embryo have an effect on the pregnancy outcome? After adjusting for confounding maternal variables, poor embryo quality was not associated with adverse obstetric or perinatal outcome in this small pilot study. Embryo quality is a major predictor of the success of in vitro fertilization treatment and studies have demonstrated a strong association between embryo morphology, implantation and clinical pregnancy rates. However, the association with obstetric and perinatal outcomes has not been evaluated. This single center, retrospective cohort study included 1541 fresh single embryo transfers (SETs) using non-donor oocytes in women ≤40 years between December 2008 and 2012. We compared the cycle outcome and singleton live births resulting from the transfer of a single fresh good quality (Grade 2) embryo with those resulting from the transfer of a single poor quality (fair, Grade 3 or poor, Grade 4) embryo in the cleavage or blastocyst stages. The cycle outcome parameters were biochemical pregnancy and clinical intrauterine pregnancy. The pregnancy outcomes were live birth, miscarriages and stillbirths after 20 weeks of gestation. Among the live births, perinatal outcome parameters included birthweight, small for gestational age, preterm delivery, pre-eclampsia, placental abruption and neonatal complications. Covariates were maternal age, body mass index, smoking status, parity and gender of the baby. There were 1193 good quality SETs and 348 poor quality embryo transfers. SETs performed during the study period resulted in 563 pregnancies and 440 singleton births. There was a higher clinical pregnancy rate (41.5%) and live birth rate (32.3%) in the good quality embryo transfer group compared with that in the poor quality transfer group (19.2 and 15.5%, respectively; P < 0.0001). There was no significant difference in the miscarriage rate between the transfers of a single good or poor quality embryo. Multivariable logistic regression analyses for pregnancy complications revealed no increased risk of maternal or neonatal complications with the transfer of a poor quality embryo. There was no difference in the obstetric or perinatal outcome of the live births resulting from a good or poor quality embryo after stratification by day of transfer. The main limitations of this study are the retrospective nature of the study, the relative subjectivity of embryo scoring and the small number of live births after transfer of poor quality embryos. Our findings may be used to reassure women that transfer of a single poor quality embryo, whether a cleavage or a blastocyst, does not appear to be associated with increased risks of adverse obstetric and perinatal outcomes. Only internal funding was used. There is no conflict of interest in relation to the study.

When looks are deceiving—the challenge facing embryo quality prognosticators

The quest for biomarkers of embryo quality is as seasoned as the practice of human ARTs itself. Reaching back into the annals of IVF reminds us of the attempts of the late Bob Edwards and his progeny during the 1980s that placed the property of fragmentation at the disposal of budding embryologists charged with identifying those embryos most suitable to transfer—and, hopefully, endowed with the most potential to give rise to a term pregnancy. As Bob showed, not so reliable was this admittedly “ugly” identifier as a predictor of implantation, let alone pregnancy. And then there were the primordial and laudable efforts of Lucinda Veeck and her colleagues to catalogue the entire range of morphological variation in oocytes, sperm, and embryos proffered to prognosticators for the next decade of human ARTs, one that brought with it measurable advances in ovarian stimulation and ICSI, placing “in the dish” many more embryos to transfer to the delight of patients and clinicians than could have been anticipated 10 years before. But, truth be told, embryo selection remains a vague and frustrating exercise, heightening the need for something less subjective and more objective even in the face of an “embryonic embarrassment of riches.” A turning point in this field of endeavor came from the pioneering efforts of the late Lynette Scott. Her passion and observational talents were harnessed through a series of papers describing the details of pronuclear development, and their attendant dynamics, providing to the ART community yet another landscape with which to discriminate between those embryos most desirable for transfer. Bearing in mind that her insights came at a time when discussions as to the optimum stage for transfer presaged the introduction of media extending the developmental window to blastocyst, pronuclear staging filled an important gap in our understanding of the earliest stages of human development. She also was among the first to deploy time-lapse imaging, convinced that cellular behaviors at the one-cell stage might someday be linked to outcome measures consistent with a term pregnancy. Now comes a report from the laboratory of Dr. Sangita Jindal taking aim at the utility of pronuclear scoring as a biomarker for pregnancy outcome (this issue JARG, Berger et al., 2014, “Embryo quality but not pronuclear score is associated with clinical pregnancy following IVF” DOI 10.1007/s10815-013-0162-3). Using more contemporary biomarkers of embryo quality such as cell number, fragmentation, and blastomere symmetry, the results clearly indicate that the application of pronuclear scoring offers no additional benefit over the use of the accepted measures cited above in terms of clinical pregnancy rates in Day 3 transfers. While putting to rest one approach to embryo scoring, and validating the use of criteria in Day 3 embryos firmly entrenched in the formative events of compaction and cavitation, this work also sets the stage for a new perspective on embryo quality assessment consonant with extending the culture period to Day 5 or 6 for embryo transfer. Why? Because along with extended culture comes the opportunity to track and deconstruct the details of individual preimplantation embryos under the watchful “eyes” of time-lapse imaging while cohorts of concepti make their merry way to the day of transfer content in their isolation in microdrops of media designed to recapitulate the female reproductive tract. It is undeniable that the introduction of dynamic imaging brings an analytical array of benchmarks to the prognosticators’ playbook. It is far less apparent to what extent the “criteria under development” will indeed offer more reliable and accurate indices of embryo quality. Here at JARG, we remain mindful of the historical traces of gamete and embryo evaluation that may at the time have satisfied the needs of embryo selection but in the end offer little more when it comes to predicting whether a clinically significant event will ever take place. While as a community of clinicians and scientists eternally optimistic that the perspective of time-lapse imaging will bear fruit as a predictive tool, “proof of the pudding” has yet to be realized. We encourage out readership to maintain focus in the pages of JARG as this next phase of embryo quality assessment matures into what will hopefully become a useful tool.

Transfer of blastocysts with deviant morphological and morphokinetic parameters at early stages of in-vitro development: a case series

Time-lapse imaging is increasingly applied as an adjunct to reproductive medicine. The gained information of the morphological and morphokinetic variables before the onset of transcription are supposed to be good predictors for the selection of the best embryo for transfer and are often seen in line with clinical outcomes. This retrospective case series investigated the outcome of transferred blastocysts that did not fulfil the proposed embryo scores at early cleavage or at later stages of development. The observations were made by time-lapse imaging. This study reports the birth of 16 healthy children after day-5 blastocyst transfer, of which at least one of the transferred embryos originated from deviant morphology and/or kinetic cleavage patterns. This case series suggests that some blastocysts derived from embryos with poor conventional morphological score and/or suboptimal morphokinetics can be successfully transferred and might result in live births. Such results might raise awareness that discarding embryos based only on early events is not a suitable approach to give patients the chance to conceive. In conclusion, to date only the transfer of viable embryos after culturing them until day 5 guarantees optimal embryo selection and helps to prevent embryo wastage.In assisted reproduction treatment, the overall success rates are still low. Optimal embryo selection is essential for IVF therapy to provide the highest chances for implantation. Therefore, several models were postulated for prediction of embryo quality. Many strategies have been applied, focusing on oocyte, zygote and early cleavage-stage morphology in a static manner. With the implementation of time-lapse technology, the information that is available in regard to embryonic development has increased tremendously. Thus, new morphokinetic markers have been established as embryo selection criteria. However, this brings the danger of wasting low-scored embryos. We here report the birth of several healthy babies after transfer of day-5 embryos that originated from oocytes or embryos that did not fulfil the morphokinetic criteria. With regard to the high costs of an IVF cycle and the emotional stress for the patient, surplus embryos should not be discarded but cryopreserved for transfer in another IVF cycle. Although the likelihood of pregnancy is lower compared to normal-scored embryos, there is still a chance of reaching blastocyst stage and successful implantation. According to our experience, the best option regarding embryo selection is still embryo culture until day 5.

Age is the Best Marker to Predict Intracytoplasmic Sperm Injection Cycles Outcome

Aims : To assess the ovarian reserve for subpopulations of Iraqi infertile couples seeking Intra Cytoplasmic Sperm Injection (ICSI) treatment. 2. To evaluate the effectiveness of ovarian reserve markers in predicting the response to Controlled Ovarian Stimulation and Intra cytoplasmic sperm injection cycles outcome. Study Design: A prospective observational controlled trial. Patients and Methods: Eighty -seven participants were enrolled during their attendance to the fertility center of Al-Sadar Medical Teaching city. Twenty subjects as a control group (N=20), and 67 as a study group (N=67). Ovarian reserve parameters were assessed and ICSI were performed. The female were divided into two groups: Those under 35 years of age and those above 35 years of age. They were compared for: Number (No.) of follicles obtained, No. of mature oocytes, Fertilization rate, Cleavage rate, Embryo scoring, No. of embryo transferred and Pregnancy rate (biochemical pregnancy). Results: The old age group has significantly higher serum follicle stimulating hormone and body mass index while they have significantly lower anti mullerian hormone than the younger age group. The ICSI outcome parameters regarding No. of oocyte, No. of mature

Embryo quality but not pronuclear score is associated with clinical pregnancy following IVF

The association between pronuclear (PN) scoring of embryos from assisted reproductive technology (ART) and clinical pregnancy remains controversial. We hypothesized that embryos with PNs scored on the day of fertilization check offer better embryo selection on day 3 and higher CPR compared to non-PN scored embryos. Patients (19-46 years) undergoing IVF/ICSI cycles at Montefiore's Institute for Reproductive Medicine and Health between January 2006 and December 2009 were included in our study. We analyzed fresh day 3 cycles only with autologous oocytes and partner's fresh sperm (n = 344). A total of 1,899 embryos were included. We compared CPR from non-PN scored embryos (Group 1, n = 835) with PN scored embryos (Group 2, n = 1,064). Composite scores by patient were developed based on embryo disposition. We also assessed traditional embryo grading derived from cell number, fragmentation and cell symmetry. Data analysis included chi square and t test to determine if PN scoring was associated with improved CPR, and to compare the additional variables. CPR between Group 1 and Group 2 were not different (p = 0.91). CPR was significantly associated with female age, number of mature oocytes retrieved, number of day 3 embryos and grade of embryos transferred on day 3 (p < 0.05). PN scoring was not associated with improved CPR in day 3 embryo transfers. Mean grade of transferred embryos continues to be a well-established, independent predictor of CPR. We conclude that further refinement of embryo grading by PN scoring is not beneficial.

Anti-Müllerian hormone levels in the follicular fluid of the preovulatory follicle: a predictor for oocyte fertilization and quality of embryo.

This prospective study investigated the relationship between anti-Müllerian hormone (AMH) level in the follicular fluid (FF) and the quality of the oocyte and embryo. A total of 65 FF samples from 54 women were included in this study. FF was collected from the largest preovulatory follicle sized≥20 mm of mean diameter from each ovary. Samples were divided into 3 groups according to the FF AMH levels: below the 33th percentile (low group, FF AMH<2.1 ng/mL, n=21), between the 33th and the 67th percentile (intermediate group, FF AMH=2.1-3.6 ng/mL, n=22), and above the 67th percentile (high group, FF AMH>3.6 ng/mL, n=22). The quality of the ensuing oocytes and embryos was evaluated by fertilization rate and embryo score. FF AMH levels correlated positively with the matched embryo score on day 3 after fertilization (r=0.331, P=0.015). The normal fertilization rate was significantly lower in the low group than in the intermediate group (61.9% vs. 95.5% vs. 77.3%, respectively, P=0.028). Our results suggest that the FF AMH level could be a predictor of the ensuing oocyte and embryo quality.Graphical

Does cigarette smoking really have detrimental effects on outcomes of IVF?

ObjectiveCigarette smoke contains many toxic chemicals associated with poor reproductive outcome and reduced fertility. It also has a negative effect on sperm motility and morphology. The aim of this study was to analyze the effects of male and female cigarette smoking on the outcomes of in vitro fertilization (IVF). Study designIn this comparative prospective analysis, the effects of smoking on outcomes of IVF including semen parameters, oocyte quality, fertilization rate, transfer day embryo scores and pregnancy rates were analyzed. For this purpose, patients were grouped based on their follicular or seminal fluid cotinine (a nicotine metabolite) levels as smokers and non-smokers (non-smokers: female (n=171), male (n=118), smokers: female (n=43), male (n=96)). ResultsThe mean age and baseline hormonal levels of all groups were found to be comparable. While the numbers of total and mature oocytes collected were higher in the smoker group (p=0.005 and p=0.006, respectively), oocyte quality index, fertilization rate, embryo development rate and pregnancy rate were not significantly different between the groups (p>0.05). Analysis based on the type of ovarian hyperstimulation protocol (GnRH agonist, antagonist and others) showed that within the antagonist group the mean age of smokers was significantly lower and the total number of collected oocytes was significantly higher compared with non-smokers. Cigarette smoking among men did not have a significant negative effect on outcomes of IVF whether their partners were smokers or nonsmokers. Regression analysis did not give any significant difference when male and/or female smoking status was analyzed for fertilization rates, transferred embryo qualities and clinical pregnancy rates. ConclusionCigarette smoking does not have detrimental effects on outcomes of IVF.

Inter- and intra-observer variability of time-lapse annotations

How consistent is the time-lapse annotation of dynamic and static morphologic parameters of embryo development, within and between observers? The assessment of dynamic parameters is characterized by almost perfect agreement within and between observers. The commonly employed method used to assess embryos in IVF treatments is based on static evaluation of morphology in a microscope, but this is limited by substantial intra- and inter-observer variation. Time-lapse imaging has been proposed as a method to refine embryo selection by adding new dynamic predictors of viability to the assessment. Yet, there are no data regarding the consistency of estimates of the time-lapse parameters. Infertile patients were recruited at the Fertility Clinic, Arhus University Hospital from February 2011 to June 2012. All embryos were cultured for 6 days in a time-lapse incubator (EmbryoScope(™)). Automated image recording was performed every 20 min. In total, 158 fertilized embryos from 20 different patients were annotated. Three observers made independent annotations on time-lapse recordings. One observer performed the assessment twice. Twenty-five parameters were annotated and the inter- and intra-observer agreement was assessed by calculating intra-class correlation coefficients (ICCs). Extremely close agreement (ICC 0.99) was found for dynamic parameters including the timing of the following: pronuclei breakdown, completion of blastocyst hatching and the appearance and disappearance of the first nucleus after the first division. Observations of cleavage divisions were strongly correlated (ICC > 0.8), indicating close agreement. Measurements of the static morphologic parameters, i.e. multi-nucleation and evenness of blastomeres at 2-cell stage showed fair-to-moderate agreement (ICC ≤ 0.5). The study was conducted at a single clinic. Only embryos with a good prognosis were included. The influence of training sessions was not measured. Consistency is crucial to the validity of embryo scoring and selection. All of the time-lapse parameters suggested by the literature showed in our study high intra- and inter-observer correlation, thus validating the precision of time-lapse annotations. This provides the basis for further investigation of embryo assessment and selection by time-lapse imaging in prospective trials. Research at the Fertility Clinic was funded by an unrestricted grant from Ferring and MSD. The authors have no competing interests to declare. NCT01139268.

Ovarian Stimulation with Urofollitropin (uFSH) Results in a Lower Yield of Oocytes Compared to Recombinant FSH (rFSH), Nevertheless, uFSH is at Least as Effective as rFSH in Younger Patients: Preliminary Results of a Retrospective Study with Antagonist Protocols in an IVF/ICSI Program

Objective: Differences in the mode of action between recombinant FSH (rFSH) and urinary derived FSH (uFSH) have been reported in cycles down-regulated by agonists. The aim of our study was to determine, if these differences also exist in cycles down-regulated by antagonists. Method: Antagonist cycles performed between 2009 - 2012 were divided into two groups: 1. Cycles stimulated with rFSH preparations: Follitropin alfa (n=203) and Follitropin beta (n=443) and 2. Cycles stimu- lated with Urofollitropin (n=405). Cetrorelix or Ganirelix were used as antagonists. All patients received 75 IU hMG additionally from day 6 of stimulation onwards up to the day of hCG administration. A logistic regression analysis was conducted to find the most significant parameters predicting hCG-positive pregnancy for 2471 IVF cycles. The results demonstrated that the predictors such as age of patients and number of cycles ever per- formed made negative contributions and number of oocytes retrieved and number of embryos transferred made positive contributions to prediction. Taking this into consideration, comparable groups could be created by including only first- ever IVF cycles with more than 10 oocytes retrieved and 2 embryos transferred. Thus, the final sample for comparison included 98 patients in the rFSH-group and 27 patients in the uFSH group. Results: There were no differences in basic personal data and gonadotropin consumption between the groups. Stimulation with rFSH resulted in a higher yield of oocytes compared to uFSH (15,6 vs. 14,4), however, the results of the following reproductive outcome parameters were all in favor of uFSH when rFSH and uFSH were compared: oocyte maturation rate (76.5% vs. 79.0 %), fertilization rate (53,6% vs. 58,6%), embryo score 4 rate (25,7% vs.. 31,1%), hCG-positive pregnancy rate (43,9% vs. 59,3%), clinical pregnancy rate (33,7% vs. 44,4%), embryo-cryopreservation rate (19,3% vs. 30,5%) and abortion rate (14,0% vs. 12,5%). Conclusions: These results seem to support the concept that uFSH produces fewer oocytes than rFSH, but the oocytes produced by uFSH are, on an average, of better quality than those produced by rFSH. Basic studies have shown that dif- ferent FSH isoforms with different elimination kinetics in the two gonadotropin preparations could be responsible for the different effects. However, the results have to be confirmed by well designed prospective randomized studies.

Long-term pituitary downregulation before frozen embryo transfer could improve pregnancy outcomes in women with adenomyosis

Some studies have shown that long-term gonadotropin-releasing hormone (GnRH) agonist administration before in vitro fertilization/intracytoplasmic sperm in infertile women with endometriosis or adenomyosis significantly increases the chances of pregnancy. We were interested in whether long-term GnRH agonist pretreatment could improve pregnancy outcomes in adenomyosis patients undergoing frozen embryo transfer (FET) after preparation of the endometrium with hormone replacement therapy (HRT). Totally, 339 patients with adenomyosis were included in this retrospective study, 194 received long-term GnRH agonist plus HRT (down-regulation + HRT) and 145 received HRT. There were no differences between the groups in characteristic such as age, body mass index, duration or cause of infertility, serum CA-125 level and basal hormone levels. On the day of progesterone administration, mean endometrial thickness and serum progesterone level were significantly greater in HRT patients. Mean score and number of embryos transferred showed no differences. In down regulation + HRT group, clinical pregnancy, implantation and ongoing pregnancy rates were 51.35%, 32.56% and 48.91%, respectively, significantly higher than that of HRT group (24.83%, 16.07% and 21.38%, respectively). So, we concluded that in FET, long-term GnRH agonist pretreatment significantly improved pregnancy outcomes in patients with adenomyosis.

The effect of the biochemical marker soluble human leukocyte antigen G on pregnancy outcome in assisted reproductive technology—a multicenter study

To determine whether the presence of soluble human leukocyte antigen G (sHLA-G) affects implantation and pregnancy outcomes in vitro. A multicenter retrospective study. Six certified in vitro fertilization (IVF) units. Embryos obtained from 2,040 patients from six different IVF clinics. Soluble HLA-G determination on day-2 embryos after intracytoplasmic sperm injection, with embryos transferred on day 3 using the sHLA-G data. Ongoing pregnancy rate (10- to 12-week ultrasound finding). All embryos were individually cultured, and a chemiluminescence enzyme-linked immunosorbent assay was used to detect the presence of sHLA-G in the culture medium surrounding the embryos. Embryos were selected based on a positive sHLA-G result and a graduated embryo scoring (GES) score >70, or on embryo morphology if the test was negative. In all centers, a positive sHLA-G result was associated with an increase in the odds of an ongoing pregnancy. The incidence of an ongoing pregnancy was 2.52 times greater in embryos transferred on day 3 with a positive sHLA-G test result than the incidence of an ongoing pregnancy in embryos with a negative sHLA-G test result. Data from this multicenter study confirm that sHLA-G expression is a valuable noninvasive embryo marker to assist in improving pregnancy outcomes, with the theoretical potential to reduce multiple pregnancies.

Analysis of the most frequented posts/comments on fertility lab insider blog reveals potential areas of improvement for patient counseling

Analysis of the most frequented posts/comments on fertility lab insider blog reveals potential areas of improvement for patient counseling

New index to asses embryo quality independently of embryonic development stage

One of the most relevant aspects in assisted reproduction is the characterization and identification of viable embryos to transfer. Many works had used pregnancy, age, number of cells at day 2 or 3 to set up scores. The purpose of this work was to analyze the predictive association of a new embryo score, contemplating number of cell; fragmentation and symmetry in embryos at day 2 and 3 in different subgroups.

Paternal age does not affect assisted reproductive outcomes when coupled with young oocytes

The objective of this study was to evaluate the effect of male age on both semen parameters and reproductive outcomes in fresh ICSI cycles with donor oocytes. Single center, retrospective cohort study of 5,089 ICSI cycles of infertile couples who elected for egg donation to overcome the female fertility issue. The association of male age with the semen parameters and laboratory outcomes were analysed by Multiple Analysis of Covariance (MANCOVA). For the laboratory outcomes the model was adjusted by semen origin (fresh vs. frozen), egg donor age and egg donor body mass index (BMI). The association of male age with the reproductive outcomes (biochemical pregnancy, ongoing pregnancy and live newborn) were modelled by logistic regression, with the following covariates: egg donor age, egg donor BMI, egg recipient age, egg recipient BMI, semen origin (fresh vs. frozen), transfer day (2 vs. 3), mean quality score of embryos transferred (<8 vs. ≥8), and number of transferred embryos (3 vs. 2 vs. 1). The adjusted odds ratios were calculated by logistic regression. Male age (41.2±6.8 y.o.) was associated with a mean decrease in seminal volume of 0.22 ml every 5 years of age (p<0.001), and with a decrease in motile spermatozoa of 1.18% for every 5 years of age (p<0.001); sperm concentration present a mean increase of 3.1 million sperm/ml (p=0.003). No significant association was found between male age and the mean transferred embryo quality (p=0.07). Male age was neither associated with biochemical (OR 1.007; CI 0.0959-1.057) or ongoing pregnancy rate (OR 0.983; CI 0.935-1.033), nor with newborn rate (OR 0.984; CI 0.936-1.035). While we confirmed the negative impact of age on various seminal parameters, we did not find any influence of paternal age on reproductive outcomes when young oocytes are used for ICSI. Further studies are needed to assess postnatal outcomes of children fathered by older men.

The effects of two different types of gondotropins on embryo kinetics

The time-lapse based system for embryo scoring is a tool for objectively evaluating the embryo development. The aim of this study was to compare the effects of two alternative regimens of controlled ovarian hyperstimulation on the kinetics of the embryo development. Retrospective study. A total of 388 embryos from 57 IVF cycles were assessed based on the time-lapse monitoring system. The inclusion criteria were age under 35, absence of azospermia or severe OAT, 1st or 2nd IVF cycle and BMI less than 25. Patients were stimulated with either human FSH-HP (Fostimon), group A, or follitropin β (Puregon), group B. In all patients the pituitary was suppressed by an antagonist protocol. The development of all the embryos obtained from these patients, 220 embryos form group A and 168 embryos from group B, was assessed using a time-lapse system (EmbryoScope). The time-lapse for cleavage to 2-cell, to 3-cell, to 4-cell, to 5-cell embryos(T2 - T5) and the duration of the division to 2 cell, to 3 cell embryo (cc2) and to a 4-cell embryo (s2) were also recorded. All the embryos were then classified in time-lapse categories from A to F as elsewhere described (Meseguer M, Hum Reprod 2011). The average age was 28.3 in both groups. The average duration of the stimulation, the mean number of oocytes retrieved and the fertilization rate were, respectively for group A and B, 9.2 vs. 9.9 days, 10.3 vs. 11.3 oocytes and 89% vs. 84% (n.s.). There were also no statistically significant differences in T2, T3, T4, T5, cc2 and s2 between embryos from both groups. The classification of embryos in time-lapse categories gave 66.4% tranferable or freezable embryos (categories A-D) in group A and 57.7% in group B, which was statistically significant. The type of gonadotropin did not affect embryo kinetics, but the classification of the embryos based on the time-lapse categories resulted in significantly higher percentage of usable embryos from the group stimulated with the human FSH-HP.

Association of serum estradiol levels on the day of hCG administration with pregnancy rates and embryo scores in fresh ICSI/ET cycles down regulated with either GnRH agonists or GnRH antagonists

To investigate the interrelation between serum E2 level on hCG day, score of transferred embryos and pregnancy achievement. Records of 350 women aged 18-40years who underwent ovarian hyperstimulation in fresh cycles down regulated either with GnRH agonist (n=70) or GnRH antagonist (n=280) followed by oocyte pick-up, ICSI and embryo transfer are retrospectively analyzed. Median E2 levels on hCG day of cycles ending with and without pregnancy were similar (p=0.308). ROC curve for AUC of E2 on hCG day with dependent variable pregnancy rate also demonstrated that the E2 level on hCG day cannot be used to predict pregnancy in IVF/ICSI cycles (AUC 0.532, 95% confidence interval: 0.471-0.593). Grouping cycles according to their E2 levels on hCG day also did not demonstrate any detrimental effect of either low or high E2 levels on hCG day both in agonist and antagonist cycles. Pregnancy rates are strongly correlated with mean and total score of transferred embryos. Interrelation of E2 on hCG day and pregnancy rate is independent from score of transferred embryos. E2 on hCG day is not correlated with pregnancy rates and cannot be used to predict pregnancy in neither agonist nor antagonist cycles, no matter its level or percentile is used.