Effect of developmental stage of embryo at freezing on pregnancy outcome of frozen-thawed embryo transfer

BACKGROUND: The study aim was to investigate the impact of the developmental stage of embryos on pregnancy outcome of frozen embryo transfer (FET). METHODS: The survival rates of embryos after thawing and pregnancy outcome following FET were compared retrospectively between three cryopreservation strategies utilizing either zygote, day 2 or day 3 embryo freezing. RESULTS: A total of 4006 embryos was analysed in 1657 thaw cycles. The highest (P < 0.0001) survival rate (all cells survived) was observed for zygotes (86.5%), followed by day 2 (61.7%) and day 3 (43.1%) embryos. FET was performed in 1586 (95.7%) of all thaw cycles, resulting in overall clinical pregnancy and implantation rates of 20.7 and 14.2% respectively. The delivery rate per transfer was 16.5%, and live birth rate per transferred embryo 11%. There were no significant differences in clinical pregnancy, implantation, delivery and birth rates between frozen zygote, day 2 and 3 embryo transfers. However, an elevated miscarriage rate was observed in the day 3 group (45%) compared with zygotes (21.3%; P = 0.049) and day 2 embryos (18.3%; P = 0.004). The overall efficacy of FET (birth rate per thawed embryo) was 7.3%. The efficacy was lower in day 3 group (4.2%) than in the zygote (7.1%; P = 0.082) and day 2 (7.6%; P = 0.027) groups. CONCLUSIONS: The developmental stage of embryos at freezing has a profound effect on their post-thaw survival, but seems to have little effect on rates of clinical pregnancy, implantation, delivery and birth after FET. The elevated miscarriage rate for day 3 frozen embryo transfers may be caused by damage during freeze-thaw procedures. The low survival rate and elevated miscarriage rate were both responsible for a reduced overall efficacy for day 3 FET when compared with zygotes and day 2 embryos.     

Embryo morphology or cleavage stage: how to select the best embryos for transfer after in-vitro fertilization

This retrospective study of 1001 in-vitro fertilization (IVF) cycles included a consecutive series of single transfers (n = 341), dual transfers (n = 410) and triple transfers (n = 250) where all the transferred embryos in each cycle were of identical quality score and identical cleavage stage. In our 2 day culture system, transfer of 4- cell embryos resulted in a significantly higher implantation rate and pregnancy rate (23 and 49%) compared with 2-cell embryos (12 and 22%) and 3-cell embryos (7 and 15%). Furthermore, the transfer of 4-cell embryos resulted in a significantly higher pregnancy rate compared with embryos that had cleaved beyond the 4-cell stage (28%). The implantation rate (21%) and pregnancy rate (43%) after transfer of embryos of score 1.0 were significantly higher than after transfer of embryos of score 2.0 (14 and 32% respectively). Transferring embryos of score 2.1 resulted in significantly higher implantation rates (26%) and similar pregnancy rates compared with score 1.0. Transferring embryos of score 2.2-3.0 resulted in a significantly lower implantation rate (5%) and pregnancy rate (15%). A striking finding was that embryos of quality score 2.0 had a significantly lower implantation rate compared with embryos of quality score 1.0 and 2.1 and a significantly lower pregnancy rate compared to embryos of quality score 1.0. We also found a lower implantation rate and pregnancy rate when transferring 3-cell embryos. These findings may indicate periods of increased sensitivity to damage during the cell cycle. In conclusion, these results substantiate the idea of the superiority of 4-cell embryos and demonstrate that minor amounts of fragments in the embryo may not be of any importance. These findings may call for a shift when weighing the two main morphological components (quality score and cleavage stage) in the sense that reaching a 4-cell cleavage stage even with the presence of a minor amount of fragments should be preferred to a 2-cell embryo with no fragments.      

Embryos cultured in vitro with multinucleated blastomeres have poor implantation potential in human in-vitro fertilization and intracytoplasmic sperm injection

The aim of the present study was to investigate pregnancy rates ensuing from transfer of embryos with multinucleated blastomeres. In our in- vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) programme, 1735 embryo transfers were performed from January 1, 1995 to August 31, 1996. In 136 of these transfers at least one embryo with one or more multinucleated blastomeres was present per transfer (study group). For each of these 136 transfers, two matched controls with transfer of exclusively mononucleated embryos were selected (control group). Matching was carried out according to age, method of fertilization (IVF or ICSI), number of transferred embryos and quality score of transferred embryos. In the study group, there were eight transfers of exclusively multinucleated embryos from which one pregnancy ensued and 128 transfers in which multinucleated and mononucleated embryos were transferred together leading to 23 pregnancies. The overall clinical pregnancy rate per transfer was 16.9% in the study group versus 28.7% in the control group (P = 0.01). The ongoing pregnancy rate per transfer was 13.2% in the study group versus 23.2% in the control group (P = 0.03). The implantation rate per transferred embryo was 6.0% in the study group versus 11.3% in the control group (P = 0.003). This study shows that embryos with one or more multinucleated blastomeres have a poorer implantation potential than embryos with mononucleated blastomeres. Transfer of embryos with multinucleated blastomeres should hence only be considered when insufficient numbers of embryos with only mononucleated blastomeres are present.      

预防OHSS行全胚冷冻的临床研究

目的探讨在辅助生育技术过程中出现卵巢过度刺激综合征(OHSS)倾向患者进行全胚冷冻,以预防OHSS发生,后进行冷冻胚胎移植的应用价值。方法选择2009年1月至2010年12月在本中心进行ART患者,共取卵1722周期,其中具有OHSS倾向患者201周期,占总取卵周期的11.67%,分极高危OHSS倾向组和高危OHSS倾向组两组。极高危OHSS倾向组给予全部胚胎冷冻,以后再安排冷冻胚胎移植,对于高危OHSS倾向组给予新鲜胚胎移植。比较促排卵药的使用总量及使用天数、HCG日E2水平、平均获卵数、平均胚胎数、平均优质胚胎数、OHSS发生率情况。比较全部胚胎冷冻进行复苏后移植与新鲜胚胎移植比较平均移植胚胎数、平均移植优质胚胎数、妊娠率、着床率、早期流产率等。结果1.两组患者的平均年龄、不孕年限、促排卵药的使用总量和天数无显著性差异;但全胚冷冻组HCG日E2水平、平均获卵数、平均胚胎数、平均优质胚胎数及OHSS发生率均高于新鲜周期移植组。2.全胚冷冻后行冷冻胚胎移植组与新鲜周期移植组的平均移植胚胎数、平均移植优质胚胎数、妊娠率、着床率、早期流产率均无显著性差异。结论 1.全胚冷冻不能减少早发型OHSS发生;2.为预防OHSS行全胎冷冻后,再行冷冻胚移植不影响不孕症患者治疗结局。     

Laser-assisted hatching increases pregnancy and implantation rates in cryopreserved embryos that were allowed to cleave in vitro after thawing: a prospective randomized study

BACKGROUND: Cryopreservation of embryos may lead to zona hardening that may compromise in vivo hatching and implantation following thawing and transfer. Assisted hatching (AH) has been advocated as a means of assisting the natural hatching process and enhancing implantation. METHODS: The aim of this study was to assess in a prospective randomized manner the effect of laser-assisted hatching (LAH) on implantation as well as clinical and multiple pregnancy rates (the primary outcome) after the transfer of frozen-thawed embryos. All embryos were thawed the day before transfer, and LAH was performed the next day on embryos that cleaved. Control group consisted of embryos that were transferred without AH. RESULTS: The performance of LAH significantly increased implantation (9.9 versus 20.1%, P < 0.01), clinical pregnancy (27.3 versus 40.9, P < 0.05) and multiple pregnancy rates (16 versus 40.3%, P < 0.07). In the LAH group, significantly more excess embryos that were left in culture hatched in vitro. CONCLUSIONS: LAH improves the outcome of frozen-thawed embryo transfer when performed before transfer on embryos that were allowed to cleave.     

The effect of intercourse on pregnancy rates during assisted human reproduction

Intercourse during an IVF cycle has the potential to improve pregnancy rates since exposure to semen is reported to promote embryo development and implantation in animals. Conversely, coitus-induced uterine contractions or introduction of infection may have a detrimental effect. A multicentre prospective randomized control trial was conducted to determine if intercourse during the peri-transfer period of an IVF cycle has any influence on pregnancy success. Participants undergoing thawed embryo transfer (Australian centre) or fresh embryo transfers (Spanish centres) were randomized either to abstain or to engage in vaginal intercourse around the time of embryo transfer. The transfer of 1343 embryos during 478 cycles of IVF resulted in 107 pregnancies (22.4%), with 125 viable embryos remaining by 6-8 weeks gestation. There was no significant difference between the intercourse and abstain groups in relation to the pregnancy rate (23.6 and 21.2% respectively), but the proportion of transferred embryos that were viable at 6-8 weeks was significantly higher in women exposed to semen compared to those who abstained (11.01 versus 7.69 viable embryos per 100 transferred embryos, P = 0.036, odds ratio 1.48, 95% confidence interval 1.01-2.19). Hence exposure to semen around the time of embryo transfer increases the likelihood of successful early embryo implantation and development.     

Zygote transfer on day 1 versus cleavage stage embryo transfer on day 3: a prospective randomized trial

BACKGROUND: It has been reported that pronuclear morphology is related to embryo quality and viability, and that zygote stage embryos might establish pregnancies after being transferred to the uterus. The objective of this study was to investigate whether transferring zygotes on day 1 would result in similar pregnancy rates compared to transferring cleavage stage embryos on day 3 in a prospective randomized trial. METHODS: Patients undergoing IVF/ICSI treatments were randomized to either day 1 or day 3 transfers by envelope withdrawal technique. Zygotes were classified as 'pattern 0' and 'non-pattern 0' according to the size and alignment of pronuclei, the number and distribution of nucleoli. The two best zygotes or embryos were transferred on day 1 or day 3 respectively. The primary outcome measure was pregnancy rate. RESULTS: Pregnancy rates were higher in day 3 group (55/131, 42%) when compared to day 1 (34/123, 28%, P = 0.024). Similarly, implantation rates were higher in day 3 group (P = 0.03). There were more cycles with cryopreservation in the day 1 group (P < 0.001). Embryo quality on day 3 was similar between pattern 0 and non-pattern 0 zygotes. CONCLUSIONS: Day 3 embryo transfers result in better pregnancy and implantation rates compared to day 1 zygote transfers. The present pronuclei scoring cannot reliably select zygotes for transfer on day 1.     

The window for embryo transfer in oocyte donation cycles depends on the duration of progesterone therapy

In 192 oocyte donation cycles performed between January 1993 and July 1996, we examined the width of 'the window for embryo transfer' using standard hormonal replacement methods. All transfers were performed within 48 h of insemination. We varied the day of embryo transfer with regard to the initiation of progesterone therapy and, thus, the duration of endometrial exposure to progesterone and analysed the resulting pregnancy rates. Patients were divided into five groups (I-V) and embryo transfers were performed 2, 3, 4, 5 or 6 days following initiation of progesterone therapy. The number of pregnancies per transfer cycle achieved in groups I-V were 0 (0%), 3 (12%), 16 (40%), 29 (48.3%), and 10 (20.4%) respectively. The increased pregnancy rate in group III in comparison to group II is statistically significant (P < 0.03). Furthermore, the pregnancy rate in group IV (5 days of progesterone administration before embryo transfer) was significantly higher than in group V (6 days of progesterone administration before embryo transfer; P < 0.005). We also noted that, when embryos were transferred 4 or 5 days after initiation of progesterone therapy, the pregnancy rates were not significantly different between menopausal and cycling recipients (50% vs 43.7%). Our results indicate that the window for embryo transfer is dependent on duration of treatment with progesterone; it begins approximately 48 h after starting progesterone administration and lasts for approximately 4 days. The optimum period for transferring embryos at the 4- to 8-cell stage corresponds to cycle days 18 and 19. Transfers performed on the 17th and 20th days of the cycle can result in successful implantation, although the rates of implantation are highest when transfers are done on days 18 and 19.      

The neglected morula/compact stage embryo transfer

BACKGROUND: This retrospective study analysed the outcomes of 339 embryo transfers on either day 3 (n = 97) or day 4 (n = 242), and proposed a grading system for morula/compact embryos. METHODS: The morula/compact embryo grading was based on: (i) the proportion of blastomeres undergoing the compaction process; (ii) the morphology of the compacted multicellular mass; (iii) the embryo quality on day 2 and 3; and (iv) the amount of fragmentation. Embryo transfers were classified into groups as follows: group I: transferred with zero 'good' embryos; group II: one 'good' embryo; group III: two or more 'good' embryos. RESULTS: Patients on day 4 were transferred with significantly fewer embryos in groups II and III (2.58 +/- 0.9 and 2.35 +/- 0.6 respectively) when compared with the correspondent day 3 transfers (3.81 +/- 1.4 and 4.07 +/- 0.9 respectively) (P < 0.05), but had the same or higher implantation and pregnancy rates. Analysing the patients who had transfers with all 'good' embryos, day 4 transfer achieved a significantly higher implantation rate compared with day 3 transfer (46.4 versus 21.4%, P < 0.01), but the number of embryos transferred on day 4 was significantly lower than day 3 (2.1 +/- 0.5 versus 3.5 +/- 0.9, P < 0.01). CONCLUSIONS: The morula/compact embryos had great value for embryo selection, which significantly reduced the number of embryos needed for transfer.     

Human embryos with unevenly sized blastomeres have lower pregnancy and implantation rates: indications for aneuploidy and multinucleation

Uneven blastomere cleavage in human embryos of 'good morphology', i.e. those normally used for transfer, is a phenomenon which has been poorly investigated. The main objective in this study was to probe deeper into the aetiology behind previous findings that embryos with uneven cell cleavage have a lower developmental capacity in comparison with evenly cleaved embryos. Our hypothesis was that uneven cleavage may result in embryos with a higher degree of aneuploidy and/or multinuclear rate, which in turn might help to explain their low implantation rate. In the first part of the study, 378 embryo transfers performed over a 3-year period were analysed retrospectively, where all the transferred embryos in each cycle were of identical morphology score and cleavage stage. In the second part of the study, multicolour fluorescence in-situ hybridization (FISH) analyses on good quality embryos, representing the uneven (n = 11) and even (n = 13) study groups were performed. When comparing day 2 transfers between 4-cell embryos, it was found that unevenly cleaved embryos had significantly lower implantation (23.9 and 36.4%) and pregnancy rates (37.6 and 52.9%) compared with evenly cleaved embryos. A significantly higher degree of aneuploidy (29.4 and 8.5%) and multinuclear rate (21.1 and 2.1%) in blastomeres from uneven embryos was also found. It is concluded that uneven blastomere cleavage has a negative effect on both pregnancy and implantation rates in human IVF, and that this can partly be explained by a higher degree of aneuploidy/multinuclear rate. In the light of the results obtained, a new approach in the current embryo scoring system, placing more emphasis on blastomere size, is recommended.     

Implantation: Two instead of three embryo transfer in in-vitro fertilization

The rational of transferring two instead of three embryos wasstudied through 468 in-vitro fertilization (IVF) treatment cyclesin 287 couples. The quality of 1224 embryos was determined accordingto the fragmentation rate and the morphology as good (A) andpoor (B). The influence of the number of embryos transferred(two or three) on the pregnancy rate when the same quality orcombinations of good and poor quality embryos transferred wasexamined. When only good quality embryos were transferred thepregnancy rates in double (AA) and triple (AAA) embryo transferwere 40.5 (17/42) and 42.9% (30/70) respectively (not significant).When only poor quality embryos were transferred, the pregnancyrates in double (BB) and triple (BBB) embryo transfers were11.0% (11/ 100) and 22.9% (16/70) respectively (P < 0.001).On the other hand, when good and poor quality embryos were transferredtogether as AB in double and as AAB and ABB in triple embryotransfer, the pregnancy rates were 36.8 (14/38) and 39.9% (59/148)respectively (not significant). There was no difference in themiscarriage rate between double and triple embryo transfers;16.7 and 18.1% respectively. The multiple pregnancy rate was14.3% for double embryo transfers and 32.4% for triple embryotransfers (P < 0.001). This study demonstrates that if thereis at least one good quality embryo available for transfer,then double instead of triple embryo transfer will not yielda significantly lower pregnancy rate. The influence of the numberof embryos transferred on the pregnancy rate became significantwhen only poor quality embryos were transferred. In conclusion,as long as at least one good quality embryo is available fortransfer, we may consider the transfer of double instead oftriple embryos.     

影响冻融胚胎临床结局的因素分析

目的探讨影响冻融胚胎移植结局的相关因素。方法收集我中心2009年570个行冻融胚胎移植周期,回顾性分析患者年龄,移植冻融胚胎质量及数量,内膜厚度对临床妊娠率、种植率、多胎率的影响。结果 Logistic回归分析表明:女方年龄、移植数、移植优质胚胎数与临床妊娠显著相关(P<0.01);多胎与女方年龄和移植数相关(P<0.01;P<0.05)。年龄<35岁的妊娠率(49.4%)明显高于≥35岁的妊娠率(37.9%)(P<0.05);移植2个胚胎的临床妊娠率(49.3%),明显高于移植1个胚胎的临床妊娠率(15.4%)(P<0.05),但与移植3个胚胎的临床妊娠率(50.0%)无差异(P>0.05);移植至少1个优胚组妊娠率(58.0%)较无优胚移植组临床妊娠率(37.5%)显著高(P<0.01)。结论女方年龄及移植的胚胎质量是影响移植后妊娠结局的关键因素。     

Selective transfer of cryopreserved human embryos with further cleavage after thawing increases delivery and implantation rates

We investigated whether further in-vitro culture of human multicellular embryos that survive cryopreservation can select the viable embryos for transfer. Embryos for cryopreservation were supernumerary multicellular embryos obtained after in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatments, with <20% of their volume filled with anucleate fragments. These had been cryopreserved using a slow-freezing and slow-thawing protocol with 1.5 M dimethylsulphoxide as the cryoprotectant. From the start of our cryopreservation programme until September 12, 1994, the thawing strategy was to thaw frozen embryos up to the exact number needed for transfer. Embryos for transfer were selected on the basis of their morphological appearance and embryo transfer to the patient was done on the day of thawing. From September 12, 1994 onwards we used a more selective thawing strategy where a cohort of up to a maximum of 12 frozen embryos per patient is thawed from which embryos of the best morphological quality, and which are furthest advanced in terms of cleavage after a 24 h in-vitro culture period in Menezo B2 medium, are selected. We took delivery rates, embryo implantation rates and birth rates into account to see if there is any difference between the following three types of transfers used: 187 transfers exclusively of embryos having continued to cleave after thawing, 107 mixed transfers of embryos with and without further cleavage and 53 transfers exclusively of embryos with no further cleavage. The overall outcome in terms of delivery rate and embryo implantation and birth rates were not different between the new and the earlier thawing policies (6.6, 5.2 and 3.6% versus 6.0, 4.1 and 2.7% respectively). Only when a distinction was made between transfers on the basis of the presence of embryos with further cleavage, did the advantage of selection on the basis of cleavage capacity become evident. Significantly higher delivery and embryo implantation and birth rates (11.2, 7.7 and 6.5% respectively) were recorded with transfers exclusively of embryos with further cleavage versus mixed transfers of embryos with and without further cleavage (1.9, 2.9 and 0.6% respectively). Fifty-three transfers exclusively of embryos with no further cleavage did not lead to any delivery. Our results demonstrate that selection of human multicellular embryos which survive cryopreservation and continue to cleave in vitro can significantly improve the delivery rate per transfer and the implantation rate per transferred embryo.      

提前复苏分裂速率较慢胚胎对妊娠率的影响

目的探讨冻融胚胎移植(FET)周期中,提前一日复苏冷冻前分裂速率较慢的胚胎对妊娠结局的影响。方法回顾性分析哈尔滨医科大学附属第一医院生殖中心2006年3月至2009年9月561例FET周期中,移植冷冻前分裂速率较慢胚胎的周期共88个。其中,33个周期为移植当日复苏胚胎,培养1-4h后移植。55个周期为移植前一日复苏胚胎,培养18-24h移植。比较两组胚胎移植后的妊娠率和种植率。结果移植当日复苏组与移植前一日复苏组妊娠率(9.1%,32.7%)与种植率(4.2%,15.5%)均有显著性差异(P﹤0.05);而患者平均年龄、移植前平均内膜厚度、胚胎复苏存活率及100%卵裂球存活率均无显著性差异(P﹥0.05)。结论对冷冻前分裂速率较慢胚胎提前一日复苏,可提高临床妊娠率和种植率。     

Parameters guiding selection of best embryos for transfer after cryopreservation: a reappraisal

BACKGROUND: The purpose of this study was to evaluate the respective influences of blastomere survival and resumption of mitosis on the outcome of frozen-thawed embryos. METHODS: A retrospective analysis was performed in our centre on 363 thawing cycles, involving 4-cell day 2 grade 1 embryos with <10% fragmentation. RESULTS: A higher implantation rate per transferred embryo was observed when all transferred embryos were characterized by fully intact blastomeres (100% blastomere survival) as compared with damaged embryos (50 or 75% blastomere survival) (22.0 versus 7.2%; P < 0.0001). Moreover, the implantation rate per transferred embryo was significantly higher for cleaved embryos compared with uncleaved embryos (19.7 versus 3%; P < 0.0001). Transfer of fully intact, cleaved embryos resulted in the highest implantation rates compared with transfer of damaged and uncleaved embryos (27.4 versus 0%; P < 0.0001). Intermediate implantation rates were observed when only one of the two criteria was fulfilled (13 versus 11% respectively; P > 0.05). Multivariate analysis showed that the clinical pregnancy rate was influenced by both criteria (odds ratio = 3.4 for transfer of embryos with six or more cells versus embryos with less than six cells. CONCLUSION: The results of our study suggest that the most important factor to predict further embryo development is the total number of blastomeres in transferred embryos, however they are obtained (good survival and/or resumption of mitosis).     

Embryo transfer: ultrasound-guided versus clinical touch.

In this prospective control study, the pregnancy and implantation rates were compared between ultrasound-guided and clinical touch uterine embryo transfers. In addition, a subset of patients was sought that would particularly benefit from embryo transfer under ultrasound guidance. A total of 187 patients (93 ultrasound and 94 clinical touch) was enrolled. Allocation was random and depended on whether their embryo transfers were done during the 1 h each day in which the ultrasound was available. Pregnancy and implantation rates of 37.8 and 20.4% respectively were achieved when ultrasound was used, compared with 28.9 and 16.2% respectively with clinical touch. This difference was not statistically significant. There was no significant difference in the pregnancy rate when the number of embryos transferred was controlled. Older women (>/=37 years old) had an apparently higher pregnancy rate (38.1 versus 20.4%; not significant) with ultrasound guidance during embryo transfer. In the subgroup where the clinician rated the transfer procedure as difficult, there appeared to be a substantial improvement in the pregnancy rate in the group that used ultrasound (54.5 versus 10.0%; not significant). Although our results were not statistically significant, we believe that ultrasound-guided embryo transfers should be used in clinically difficult embryo transfers and in older women, as it appears to improve the pregnancy rate over clinical touch transfers.     

Cryo-thawed embryos obtained from conception cycles have double the implantation and pregnancy potential of those from unsuccessful cycles

BACKGROUND: The purpose of this study was to evaluate the influence of fresh IVF/ICSI cycle outcome on the prognosis of the related frozen embryo replacement (FER) cycle. METHODS: 459 FER cycles, involving 2049 cleavage stage embryos with no or up to 10% fragmentation, were performed for which the outcome of the fresh cycle was recorded. The cycles were divided into two groups; group A included cycles in which cryopreserved embryos were obtained from fresh cycles in which conception occurred. Group B were cycles in which cryopreserved embryos originated from unsuccessful fresh cycles. RESULTS: Groups A and B were comparable with respect to mean (+/- SD) age at cryopreservation (33 +/- 3.9 versus 33.2 +/- 4 years, P = not significant), mean number of oocytes retrieved and fertilized normally in the fresh cycle (11 +/- 5.2 versus 11.2 +/- 4.8, P = not significant) and mean age at the cryo-thawed transfer (34.5 +/- 4.2 versus 33.9 +/- 4 years, P = not significant). No significant difference was found between the two groups with regard to mean number of embryos cryopreserved (6.5 +/- 3.9 versus 6.2 +/- 3.6) and subsequently thawed (4.5 +/- 2.5 versus 4.5 +/- 1.8) per cycle and number of cryo-thawed embryos transferred per cycle (2.0 +/- 0.7 versus 2.1 +/- 0.8). However, the implantation rate per transferred embryo in group A was double that in group B (23 versus 11.2%, P < 0.0001). Moreover, the clinical pregnancy and ongoing pregnancy rates per cycle were significantly higher in group A compared with group B (34.8 and 27.3% versus 15.6 and 13.1%, P < 0.0001 and P = 0.0003 respectively). The difference in FER cycle outcome could not be explained by confounding variables. CONCLUSIONS: After thawing, cryopreserved embryos originating from conception IVF/ICSI cycles achieve double the implantation and pregnancy rates of those obtained from unsuccessful cycles.     

A prospective, randomized study comparing day 2 and day 3 embryo transfer in human IVF

It is believed that delayed transfer of embryos after IVF allows for a better selection of good quality embryos. Hence, the number of embryos and all other prognostic factors being equal, transfer of day 3 embryos should be associated with higher implantation and pregnancy rates than transfer of day 2 embryos. To investigate this hypothesis, a prospective randomized study was carried out to compare implantation and pregnancy rates between day 2 and day 3 transfers. The relationship between the embryo quality score of day 2 and day 3 embryos and their respective implantation rates was also analysed. In a 2 year period all patients undergoing infertility treatment and in whom at least seven normally fertilized oocytes were obtained were included in the study. A minimization procedure was performed taking into account the patient's age and the method of fertilization (IVF or intracytoplasmic sperm injection). By using a uniform policy of embryo transfer, the number of embryos transferred was similar in both groups. The outcome parameters were embryo quality, implantation and pregnancy rates. No difference was observed in implantation and pregnancy rates between transfers on day 2 versus day 3 (23.8 versus 23.8% and 47.9 versus 46.8% respectively). The incidence of embryos of moderate to poor quality was higher in embryos cultured for 3 days compared with those cultured for 2 days. It is concluded that the outcomes of embryo transfer in terms of implantation and pregnancy rates are comparable for day 2 and day 3 embryos, although the overall embryo quality score decreases when embryos are kept in culture till day 3.     

Implantation: Embryo score to predict implantation after in-vitro fertilization: based on 957 single embryo transfers

The purpose of this study was to devise an embryo score to predictthe likelihood of successful implantation after in-vitro fertilization(IVF). Unlike most studies dealing with the influence of embryostage and morphology on pregnancy, our study was based on singlerather than multiple embryo transfers. A total of 957 singleembryo transfers were carried out. No delivery was obtainedafter any of the 99 transfers using 1-cell embryos or embryosobtained after delayed fertilization. In the remaining 858 transfers,the embryos had cleaved. Higher pregnancy rates were obtainedwith embryos displaying no irregular cells (11.7 versus 6.9%;P < 0.01) and embryos displaying no fragmentation (11.5 versus8.1%; P < 0.05). The 4-cell embryos implanted 2-fold moreoften than embryos with more or less cells (15.6 versus 7.4%;P < 0.01). Based on these observations, we devised a 4-pointembryo score in which embryos are assigned 1 point each if they(i) are cleaved, (ii) present no fragmentation, (iii) displayno irregularities, and (iv) have four cells. Both pregnancyrate and take home baby rate were significantly correlated withembryo score. Each point of this score corresponds to a 4% increasein pregnancy rate. Interestingly, pregnancy rate was significantlylower in women aged >38 years (8.2 versus 11.4%; P < 0.05),even though embryo quality was similar regardless of age. Singleembryo transfer allowed us to define a simple and useful embryoscore to choose the best embryo for transfer to optimize IVFand embryo transfer outcome. The use of this embryo score coulddecrease multiple pregnancies after multiple embryo transfers.     

Impact of patients' choice for single embryo transfer of a top quality embryo versus double embryo transfer in the first IVF/ICSI cycle

BACKGROUND: The aim of this study was to evaluate the impact of transferring a single top quality embryo in the first IVF/ICSI cycle of patients <38 years old who chose to have one or two embryos transferred. METHODS: A total of 262 patients participated in the study, and 243 transfers were performed: 156 (64%) patients chose the transfer of a single top quality embryo, if available, and two non-top quality embryos if not available; 87 (36%) patients chose to have a double embryo transfer regardless of embryo quality. RESULTS: In the first group an ongoing pregnancy rate of 40% (63/156) with a twin pregnancy rate of 2% (1/63) was achieved. In the second group the ongoing pregnancy rate was 44% (38/87) with 26% (10/38) twin pregnancies. In the patient group with only one embryo transferred, irrespective of the patient's choice, the ongoing pregnancy rate was 43% (54/127) with no twin pregnancies. For the study population as a whole, the ongoing pregnancy rate was 42% (101/243) with 11% (11/101) twins. CONCLUSION: We conclude that the introduction of single embryo transfer in the first IVF/ICSI cycle is highly acceptable in women <38 years old.