Embryo Score Research Articles

Graduated Embryo Score and soluble human leukocyte antigen-G expression improve assisted reproductive technology outcomes and suggest a basis for elective single-embryo transfer

To evaluate assisted reproductive technology (ART) outcomes by using Graduated Embryo Score (GES) and soluble human leukocyte antigen-G (sHLA-G) expression to select embryos for transfer on day 3. Prospective cohort. Private practice. Women undergoing fresh ART cycles (n = 209). In vitro fertilization using standard protocols. Embryos scoring GES of > or =70 using were selected for transfer on the basis of sHLA-G expression in the culture media on day 2. Pregnancy, implantation, and multiple-gestation rates. Ongoing gestations increased with the number of embryos expressing sHLA-G (37%, 42%, 58%, and 56% with 0, 1, 2, or 3 sHLA-G(+), respectively). With at least two sHLA-G(+) embryos, ongoing gestation and implantation rates were higher than those with fewer than two sHLA-G(+). Differences were even higher for women aged < or =37 years. With at least two sHLA-G(+) embryos, the odds ratio (95% confidence interval) was 1.59 (1.51-1.68) for ongoing gestation compared with the case of fewer than two sHLA-G(+). Age was the most important predictor of outcome; the odds ratio (95% confidence interval) was 2.07 (1.98-2.16) for ongoing gestation in women aged < or =37 years with at least two sHLA-G(+) embryos, compared with the case of women aged 38-40 years. Day 3 embryo transfer using GES and sHLA-G improves ART outcomes by increasing predictive accuracy. High twin rates suggest that couples with at least two sHLA-G(+) embryos consider elective single-embryo transfer.

Relationship between even early cleavage and day 2 embryo score and assessment of their predictive value for pregnancy

The aim of this study was, firstly, to define the different patterns of early cleavage (EC) observed at 26 h after either IVF or intracytoplasmic sperm injection (ICSI) and, secondly, to assess the predictive value of one of these patterns, even EC (EEC), on pregnancy rate in combination with day 2 embryo score. In the first part of the study, the relationship between three different EC patterns (EEC, uneven EC and EC with fragmentation of the day 2 embryo) and embryo morphology was determined. EEC was shown to be strongly associated with good embryo morphology. In the second part of the study, it was shown that EEC used in combination with embryo score improved selection of embryos for transfer. The presence of EEC significantly ( P < 0.001) enhanced mean implantation rate in all transfer categories involving identically scored embryos, in both compulsory single embryo transfers and elective single embryo transfers. Multivariate analysis demonstrated that EEC and embryo score had strong complementary predictive value for pregnancy. Based on these findings, it was concluded that even though they are associated, EEC and embryo score could be combined to increase pregnancy rate, especially in elective single embryo transfer programmes.

Construction of an evidence-based integrated morphology cleavage embryo score for implantation potential of embryos scored and transferred on day 2 after oocyte retrieval

Evidence-based morphological embryo scoring models for ranking of implantation potential are still scarce, and the need for a precise model increases when aiming for singleton pregnancies. Prospectively, 2266 IVF/ICSI double-embryo, day 2 transfers were studied. The five variables scored in 3- to 5-step scales for the embryos transferred are blastomere number (BL), fragmentation, blastomere size variation ('equality', EQ), symmetry of the cleavage and mononuclearity in the blastomeres (NU). The scoring results of embryos with an individual traceability from scoring to implantation, i.e. treatments resulting in either no implantation (n=1385) or twin implantation (n=228), were studied for prognostic potential. Although all five variables correlated highly with implantation potential, only BL, NU and EQ remained independently significant after regression analysis. The equation thus derived formed the basis for a 10-point integrated morphology cleavage (IMC) embryo score. A table with the scoring point for each possible combination of the embryo variables is presented. The scoring model was statistically validated on the singleton pregnancy group (n=653). We suggest that this IMC embryo scoring, incorporating cleavage stage and information on the variation in blastomere size and the number of mononucleated blastomeres, may optimize embryo ranking and selection for day 2 transfers.

Effect of reduced oxygen concentrations on the outcome of in vitro fertilization

We compared the effects of two standard oxygen concentrations, physiological (5% O(2), 5% CO(2), and 90% N(2)) and atmospheric (5% CO(2) with the balance as air), on fertilization, embryo development, and pregnancy rate in 106 patients undergoing IVF, excluding donor oocyte cycles and preimplantation genetic diagnosis cycles. The differences in oxygen concentration did not significantly affect fertilization rate, blastocyst formation, or pregnancy rate, but there was a significant difference in mean embryo score between physiological and atmospheric groups on day 3.

Insulin-like growth factor-II (IGF-II), IGF-binding protein-3 (IGFBP-3), and IGFBP-4 in follicular fluid are associated with oocyte maturation and embryo development

The objective of this study was to investigate the association between follicular fluid (FF) levels of insulin-like growth factors (IGFs), IGF binding proteins (IGFBPs), and pregnancy-associated plasma protein-A (PAPP-A), which is a protease for IGFBP-4, and the quality of subsequent embryo development from in vitro fertilized oocytes aspirated from the same follicle. Prospective study. University infertility clinic and academic research laboratory. One hundred sixty-two infertile women undergoing controlled ovarian hyperstimulation for IVF and embryo transfer were recruited in a university hospital. During oocyte retrieval, samples of 225 FFs and matched mature oocytes were collected and studied. Concentrations of FF IGF-I, IGF-II, IGFBP-1, IGFBP-3, IGFBP-4, and PAPP-A were determined using ELISA. Progesterone secretion by cultured granulosa cells (GC) was measured by RIA. Levels of IGF-II, IGFBP-3, and IGFBP-4 in FF on the day of oocyte retrieval were significantly correlated with embryo scores on day 3 (72 hours after oocyte retrieval). Levels of IGF-II, IGFBP-3, and IGFBP-4 in FF from follicles in which oocytes developed into day 2 embryos (48 hours after oocyte retrieval; 20 >or= embryo score >or= 6) after fertilization were significantly higher than those from follicles in which oocytes were unable to be fertilized and were arrested in embryo development on day 2, whereas the levels of PAPP-A were significantly lower in the former than the latter group. Using multiple regression analysis, we found that high levels of IGFBP-3 and IGFBP-4 combined with low levels of PAPP-A in FF were significantly correlated with successful fertilization and early development into day 2 embryos. In contrast, high FF IGFBP-1 and IGFBP-4 in combination with low FF IGF-I were significantly correlated with a later (day 2-day 3) embryo development. A significant stimulation of P secretion in cultured GCs by the combination of recombinant IGF-II, IGFBP-3, and IGFBP-4 further strengthened these proteins' functional roles in promoting late follicular development. High IGF-II, IGFBP-3, IGFBP-4, and low PAPP-A levels in FF at the time of oocyte retrieval suggest better oocyte maturation and early embryo development (within 48 hours after oocyte retrieval), whereas high IGFBP-1, IGFBP-4, and low FF IGF-I levels may favor later embryo development (between 48 and 72 hours after oocyte retrieval).

Prediction of embryo developmental potential and pregnancy based on early stage morphological characteristics

To analyze the association between morphological details at different stages of culture with blastocyst development, with an aim to improve selection for transfer. Retrospective audit of data. Tertiary referral center and university hospital. Two hundred sixty-eight couples underwent 357 treatment cycles. Oocyte pickups for IVF or intracytoplasmic sperm injection (ICSI) after ovarian stimulation. Embryos were individually cultured and examined on days 0-2 for morphological details and developmental characteristics, and selected for transfer, freezing, or further culture. The association of blastocyst development and pregnancy with morphological characteristics. Five morphological characteristics (appearance of the cytoplasm, pronuclei and nucleoli, cytoplasmic deficit, and developmental rate) showed the strongest association with blastocyst development. By combining information from all days of culture into a cumulative score, prediction was greatly improved, compared to only using day 2 morphology. Cytoplasmic dysmorphisms of the oocyte, including accumulation of smooth endoplasmic reticulum, were associated with poor developmental performance. Differential weighting of these characteristics was included in a new embryo scoring system, which showed a strong correlation with implantation. Weighting individual morphological characteristics of zygotes and embryos and combining them into a cumulative embryo score can improve selection of embryos for transfer.

Effect of dehydroepiandrosterone on oocyte and embryo yields, embryo grade and cell number in IVF

The aim of this study was to investigate the effect of treatment with dehydroepiandrosterone (DHEA) on fertility outcomes among women with diminished ovarian reserve. This is a case-control study in an academically affiliated private infertility centre. Twenty-five women with significantly diminished ovarian reserve had one IVF cycle before and after DHEA treatment, with otherwise identical hormonal stimulation. Women received 75 mg of DHEA daily (25 mg three times daily) for an average of 17.6 +/- 2.13 weeks. We performed a comparison of IVF outcome parameters, before and after DHEA treatment, including peak estradiol (E(2)) levels, oocyte and embryo numbers, oocyte and embryo quality and embryo transfer statistics. Paired analysis of IVF cycle outcomes in 25 patients, who underwent cycles both before and after DHEA supplementation, demonstrated significant increases in fertilized oocytes (P < 0.001), normal day 3 embryos (P = 0.001), embryos transferred (P = 0.005) and average embryo scores per oocyte (P < 0.001) after DHEA treatment. This study confirms the previously reported beneficial effects of DHEA supplementation on ovarian function in women with diminished ovarian reserve.

Factors determining early pregnancy loss in singleton and multiple implantations

The incidence of first trimester pregnancy loss is much lower in IVF twin pregnancies than in IVF singleton pregnancies. The objective of this study was to determine which embryonic and maternal factors contribute to this finding. Retrospective data analysis of the outcome of 1593 pregnancies after day 3 double-embryo transfer (DET) after IVF or ICSI treatment. Of 1148 single implantations at 6 weeks, 936 (81.5%) were ongoing pregnancies. Of 445 multiple implantations at 6 weeks, 354 (79.6%) were ongoing multiple pregnancies, 80 (17.9%) were ongoing singleton pregnancies and 11 (2.5%) ended in a spontaneous abortion. Total pregnancy loss was 18.5 and 2.5% (P < 0.001) in singleton and twin gestations, respectively. Loss per gestational sac was 18.5 and 11.46% (P < 0.001), respectively. Determinants contributing to the continuation of gestation beyond 6 weeks were young maternal age, possibility to cryopreserve embryos and short GnRH agonist flare-up stimulation protocol. Whereas factors promoting multiple implantation at 6 weeks of gestation were young maternal age, high cumulative embryo score (CES), male infertility, long stimulation protocol and thick endometrium. Although multiple implantation at 6 weeks is predominantly determined by (morphological) embryo quality, the continuation of pregnancy beyond 6 weeks becomes more dependent on the combination of genetic and developmental potential of the embryo(s) and an optimal uterine milieu.

P-120: A prospective randomized pilot study comparing pregnancy rates and implantation rates when embryos are selected based on a single day 3 (D.3) score vs. a graduated embryo score (GES) plus soluble human leukocyte antigen-G (sHLA-G) expression

In most IVF programs, the morphological characteristics of the embryos are evaluated on the day of transfer and the embryos with the highest morphological scores are transferred. We compared two different embryo-scoring methods, D.3 vs. GES and found no differences in either the pregnancy rates or the implantation rates with either scoring method. Embryos expressing a high concentration of sHLA-G, have been shown to improve implantation rates. The aim of this study was to compare pregnancy rates and implantation rates when embryos are selected based on a single D.3 score vs.

O-157: Higher on-going pregnancy rate from day 3 frozen embryo transfer than from frozen blastocyst transfer when embryos are cryopreserved on day 3 based on graduated embryo score (GES): A randomized trial

O-157: Higher on-going pregnancy rate from day 3 frozen embryo transfer than from frozen blastocyst transfer when embryos are cryopreserved on day 3 based on graduated embryo score (GES): A randomized trial

P-997: Clinical use of pentoxifylline for freeze-thawed spermatozoa not improve embryo quality

Pentoxifylline (PTX) is a metilxantina, clinically used as an artificial sperm stimulant to enhance sperm motility in many in vitro fertilization (IVF) programs. By inhibiting phosphodiesterase, pentoxifylline increases the intracellular cAMP level, and thus contributes to sperm capacitation, hyperactivation, and acrosome reaction. With the introduction of ICSI, its evident the ability to achieve high fertilization and pregnancy rates regardless semen parameters. Moreover, the use of cryopreserved sperm was then developed with success, and sperm motility after freeze-thawed procedure is one of the main determinants of the successes of ICSI. This study was designed to evaluate the effect of PTX applied to freeze-thawed spermatozoa, on fertilization rates and embryo development after intracytoplasmic sperm injection (ICSI) cycles. Prospective experimental study. This pilot study included 15 couples who underwent assisted reproduction techniques (ART) by ICSI using frozen-thawed spermatozoa. After thawed, the sperm samples were processed by swim-up, and divided into two equal aliquots. The first was treated with 5mM PTX solution (PTX group) for 30 minutes at 37°C, and other without PTX (control - CT group), followed by sperm-wash and motility evaluation. The sperm samples were addressed to IVF laboratory for ICSI, and 117 oocytes in metaphase II (MII) were injected, 58 with spermatozoa of CT group, and 59 with PTX treated sperm. The fertilization rate, pro-nuclei morphology, early cleavage and embryo morphology on day +3 (transfer day) were evaluated. Mann-Whitney and qui-square testes were used as appropriate, p < 0.05 were considered statistically significant. Our results showed that spermatozoa treated with PTX presented total motility similar to CT group (30.1% ± 16.4 versus 34.9% ± 16.8, respectively; p = 0.534), as soon as the progressive motility (19.1% ± 14.2 versus 24.1% ± 15.7, respectively; p = 0.383). After ICSI, the normal fertilization rates, were 78.4% in the PTX group and 78.9% in CT (p = 0,456), and resulted in 82 embryos in which morphology parameters were evaluated. The percentage of embryos presenting good pro-nuclei morphology (PTX: 22.5% versus CT: 18.4 %; p = 0.655), as well as early cleavage (PTX: 32.1% versus CT: 28.6%; p = 0.838) were similar. Also, 58.1% of embryos on PTX group and 80.7% on CT, presented good morphology on day +3 (p = 0.097). A cumulative embryo score, according to morphological embryo parameters since fertilization day until transfer day were determined, and on the PTX group 58.6% of the embryos were considered good, while 82.8% in CT were in this class (p = 0.082). Embryos originated from ICSI using spermatozoa from both PTX and CT groups were transferred, and a total of 5 pregnancies were achieved (pregnancy rate: 36%). Although those are preliminary results, it was observed that the treatment of spermatozoa with PTX, in comparison with nontreated group, had similar results neither to improve the total and progressive sperm motility, nor to the other embryo parameters evaluated. It is known that the pentoxifylline is embryo toxic; indeed we found that although the sperm characteristics were similar, the embryo quality seems to be worse when the pentoxifylline was applied, suggesting that this protocol is not efficient. We propose that other protocols can be used in order to improve the motility of thawed spermatozoa and the embryo quality.

P-14: Soluble HLA-G (sHLA-G) expression results in improved ART outcomes from transfer of morpholoogically normal embryos with a high graduated embryo score (GES): A move toward single embryo transfer

P-14: Soluble HLA-G (sHLA-G) expression results in improved ART outcomes from transfer of morpholoogically normal embryos with a high graduated embryo score (GES): A move toward single embryo transfer

Embryo quality is more important for younger women whereas age is more important for older women with regard to in vitro fertilization outcome and multiple pregnancy

To determine the efficiency of embryo scoring systems for multiple pregnancy in women undergoing IVF procedures. Retrospective record analysis. University hospital, tertiary medical center. Three hundred one patients undergoing controlled ovarian stimulation, IVF/intracytoplasmic sperm injection (ICSI), and day 3 embryo transfer. IVF/ICSI and embryo transfer. Rate of pregnancy and rate of multiple pregnancy. The score of the best three embryos (Top3) was more correlated with IVF outcome than were the number of good embryos (P=.009) or the cumulative embryo score (P=.038). In the logistic regression model, Top3 was more relevant to IVF outcome and multiple pregnancy for younger patients than was age (P<.05). For older patients, age was more correlated with IVF outcome and multiple pregnancy than was embryo morphology (P<.05). The embryo morphology criteria can help reduce the number of embryos transferred into younger patients. We could use the age of patients as an indicator to determine the number of embryos transferred into older patients.

Transfert électif d'un seul embryon : une option justifiée pour une population de patientes sélectionnées

Prevention of twin pregnancies using elective Single Embryo Transfer (e-SET) is now considered by many Assisted Reproductive Techniques teams as a necessity. The aim of this study was to assess the efficacy of e-SET in a prospective manner in a selective population of patients using Take Home Baby Rate per couple as principal parameter. This prospective study was conducted from January 2003 to December 2004. Elective Single Embryo was proposed to women above 37 years in their first IVF or ICSI attempt. It was then performed only in cases when at least one embryo with high implantation potential (score-4 embryo in our embryo scoring) was obtained for transfer and one more (score-3 or score-4 embryo) was available for freezing. e-SET was proposed and accepted in 225 couples (25% of eligible couples and 7.8% of total population) and was possible in 96 of these). Two embryos were transferred in all other eligible patients (Double Embryo Transfer group=DET). Cumulative delivery rate after fresh embryo transfers and, if necessary, after frozen-thawed embryo transfers were 39.5% per couple e-SET group and 41.7% in DET group (NS). On the other hand, the percentage of twin pregnancies was significantly different between the two groups (2.6% vs 26.6% respectively; P<0.01). In women younger than 37 years in their first IVF/ICSI attempt, the elective transfer of only one embryo with high implantation potential strongly allowed to avoid twin pregnancies without any significant delivery rate decrease. This transfer policy is particularly efficient in laboratories displaying good results in their embryo freezing program.

Assessment of the ovarian volume, number and volume of follicles and ovarian vascularity by three-dimensional ultrasonography and power Doppler angiography on the HCG day to predict the outcome in IVF/ICSI cycles

The aim of this prospective study was to investigate whether ovarian blood flow is related to embryological parameters and whether it could be a predictor of outcomes of IVF/ICSI. Eighty infertile women underwent ovarian stimulation with gonadotrophins after a long protocol with GnRH agonists. The ovarian volume (OV), number of follicles (NF) and follicular volume (FV) of all follicles >10 mm and vascularization index (VI), flow index (FI) and vascularization-flow index (VFI) were obtained by three-dimensional (3D) ultrasonography and power Doppler angiography (PDA) on the day of HCG administration. These parameters were tested for their relation with IVF laboratory parameters. The OV, FV, VI, FI and VFI were significantly greater in the pregnant group. The NF and FV were the only independent predictors of the number of oocytes retrieved, mature and fertilized, and the number of embryos developed and their cumulative embryo score. Nevertheless, the number of grade 1 embryos depends on the NF and the VI. The ovarian FI and the number of transferred grade 1 embryos can predict gestation in 76% of IVF patients. A low FI and non-grade 1 embryo transferred are also associated with an increased pregnancy loss. 3D ultrasonography and PDA allow for an easier ovarian assessment in IVF cycles. The predictive value of IVF outcome suggests a high clinical usefulness of this new technique.

Impact of the assessment of early cleavage in a single embryo transfer policy

The policy of single embryo transfer (SET) adopted for women <36 years old since 1 July 2003, strongly calls for improvement of embryo selection. A total of 196 cycles in which SET was performed were randomly allocated to two groups. In the first group, early cleavage was assessed (ECA) 25 h after insemination. The embryo with the best score that cleaved early, if present, was selected for transfer. In the second group, early cleavage was not assessed (ECNA) and embryo selection was based solely on the embryo score. Ninety-eight cycles were allocated in the ECA and ECNA group respectively. Early cleavage occurred in 64% of cycles and 32.2% of embryos. Patient population and embryo morphology were similar between the two groups, and similar delivery rates were observed (27.6 versus 24.5% respectively in the ECA and ECNA groups). The assessment of early cleavage as additional parameter did not improve the delivery rate in the single embryo transfer policy.

Embryo Cleavage Itself is a Strong Predictor of Embryo Quality and in Human IVF

Selecting the right embryos for transfer in assisted reproductive technology (ART) seems to be very important in terms of outcome; although this selection is not easy to do. Amongst other strategies, many embryo scores have been proposed for embryo selection in order to achieve a higher implantation and pregnancy rate, while minimizing the risk of multiple pregnancy. However, there is no consensus about which the best parameter is. Generally, scores include morphological criteria such as: cell fragmentation, blastomere and embryo symmetry, and the use of complex formulas to make an appropriate score. Many authors suggest that the fastest cleaving embryos are considered to have the best developmental potential acting as an individual prognostic factor. The objective of this study was to analyze whether the cleavage rate is a better predictor of implantation and pregnancy rate than the embryo morphological score. None A cohort of 769 cycles of women undergoing their first fresh embryo transfer (ET) was prospectively evaluated. Inclusion criteria were: age 18 to 39 years, basal FSH levels ≤ 12 IU/L, and 3 or 4 embryos transferred to the uterus on day 3. Oocyte donation cycles were excluded. Embryos were scored considering: cleavage rate, and embryo morphology according to cell fragmentation (%), and embryo and blastomere symmetry (embryos were scored 1 to 4, being 4 the best). Global embryo score (GES) was calculated by adding the score of every single embryo transferred. Best quality embryos added a GES of 12 points or above. Cleavage rate was calculated by adding the number of blastomeres of all the embryos considering a high cleavage rate when it was equal to or above 24 points. Statistical analysis was made using chi2 test for proportions, and t-test for continuous variables. Mean age of the study group was 33.6 ± 3.6 years, and ages between pregnant and non pregnant patients was comparable.Clinical pregnancy rate was 37.1 %. All pregnancy cycles had a GES of 9.5 ± 3.5 and a cleavage rate of 19.1 ± 7.5. Women who got pregnant had a GES 2 points higher and a cleavage rate 4 points higher than cycles without pregnancy (p<0.001).In terms of pregnancy achievement potential, good quality embryos had a sensitivity of 23.2%, with a specificity of 84.7%. When only cleavage rate was considered; 31.2% of women who got pregnant had high cleavage rates, while 82.8% of those who did not get pregnant had low cleavage rates (p<0.05) Although no good accurate pregnancy prognosis scores have been developed, many variables are known to be related with ART outcome. Whenever variables are simpler, its interpretation and use are easier. In this study, cleavage rate alone showed not only to be at least as specific as the morphological score requiring three components, but it was also more sensitive. We consider that this is not a minor finding when analysing which embryo is to be transferred to maximize results at low risks; and for research when we wonder which variables have to be considered to make a more accurate prognosis.

Influence of Embryo Scoring and Number of Embryos Transferred on Clinical Pregnancy Outcomes in ICSI Cycles

Embryo quality and number have been thought to be important factors involved in optimizing outcome in human IVF. Transferring 3 embryos is a routine practice at our center. However, recently we have begun to transfer 2 embryos on informed consent. Embryos are scored from 1 (poor) to 5 (excellent) with a 5-point scoring system at our center. Embryos of score 3 to 5 are rendered transferable. This paper aims to evaluate the influence of these two factors on pregnancy rate in ICSI cycles.

Coasting Decreases Ovarian Hyperstimulation Syndrome in Patients at Risk. Is the Quality of Oocytes, Embryos and Pregnancy Rate the Price to Pay?

To determine the impact of coasting and hCG dose on the incidence of ovarian hyperstimulation syndrome (OHSS), clinical pregnancy rate, fertilization rate, and cumulative embryo score in hyper-responsive patients compared with normo-responsive patients.

Peculiar.. Embryo Phenotypes, Are They Related With Embryo Quality, Do They Affect IVF Outcome?

Classical human embryo score normally takes into account the number of blastomeres, percentage and type of fragmentation, multinucleation, symmetry and compaction. However, other features such as presence of vacuoles (VAC), smooth endoplasmic reticulum (SER) and blastomeres with cytoplasm polarization (POL), are also present in human embryos. Furthermore, high scored embryos using the classical system may also show these phenotypes. The objective of this study is to analyze whether the appearance of these unusual phenotypes affect further embryo development and the IVF outcome of the cycle