Higher clinical pregnancy rates from frozen-thawed blastocyst transfers compared to fresh blastocyst transfers: a retrospective matched-cohort study

Purpose

The purpose of this matched-cohort study was to assess endometrial receptivity to blastocyst implantation in fresh embryo transfer by comparing implantation outcomes of fresh embryo transfer with frozen embryo transfer, where two blastocysts of good quality were transferred in good prognosis patients.

Method(s)

Fresh embryo transfer from intracytoplasmic sperm injection cycles and artificial frozen embryo transfer cycles performed from January 2012 to December 2013 at a private clinic were retrospectively analyzed and the pregnancy, clinical pregnancy, and implantation rates statistically compared. Cycles were stratified and matched according to the blastocyst expansion grade (grade 2, 3, 4, or 5) of the two blastocysts transferred. Five hundred ninety-eight fresh embryo transfer cycles were matched with 545 frozen embryo transfer cycles across four blastocyst cohorts.

Result(s)

In this study of 1143 blastocyst transfer cycles, fresh embryo transfer resulted in reduced pregnancy, clinical pregnancy, and fetal heart implantation rates in all four blastocyst cohorts. The fetal heart implantation rates for fresh embryo transfer ranged from 16.7 % in the grade 2 to 47.3 % in the grade 5 cohort, compared to 23.1 % in the grade 2 to 57.4 % in the grade 5 cohort for frozen embryo transfer. The trends in increasing pregnancy outcomes relative to increasing blastocyst expansion were similar in fresh embryo transfer and frozen embryo transfer.

Conclusion(s)

Blastocysts of good quality transferred in frozen embryo transfer had a significantly greater chance of implantation and clinical pregnancy compared to blastocysts of matched quality transferred in fresh embryo transfer, suggesting reduced endometrial receptivity in fresh embryo transfer.     

Effect of the site of assisted hatching on vitrified-warmed blastocyst transfer cycles: a prospective randomized study

Purpose

To assess the effect of assisted hatching (AH) site on the clinical outcomes in vitrified-warmed blastocyst transfer cycles.

Methods

A total of 160 women who underwent vitrified-warmed blastocyst transfer cycles were randomized to either the ICM group (AH performing at the site near the inner cell mess, ICM), or the TE group (AH performing at the site opposite to the ICM). AH with laser zona drilling was performed 20 or 30 min after thawing once the ICM can be detected. Clinical pregnancy rate, implantation rate, live birth rate and the occurrence rate of monozygotic twins (MZT) pregnancy after transfer of these two groups were compared.

Results

No significant difference was found in the clinical pregnancy rate (63.8 % vs. 67.5 %), implantation rate (51.7 % vs. 53.6 %) and live birth rate (57.5 % vs. 62.5 %) between the ICM group and the TE group. The occurrence rate of MZT was comparable between the two groups (3.9 % vs. 5.6 %).

Conclusions

The site of assisted hatching has no influence on the implantation, pregnancy and live birth rate in human vitrified-warmed blastocyst transfer cycles.     

Use of the natural cycle and vitrification thawed blastocyst transfer results in better in-vitro fertilization outcomes

Purpose  

To compare the IVF outcomes of vitrification-thawed blastocyst transfer cycles utilizing different endometrial preparation methods.     

Comparison of clinical outcomes between single and double vitrified-warmed blastocyst embryo transfer according to the day of vitrification

Purpose

To compare the efficacy of single vitrified-warmed blastocyst embryo transfer (SVBT) versus double vitrified-warmed blastocyst embryo transfer (DVBT) according to the day of vitrification.

Methods

This retrospective study included a total of 1,051 cycles in women less than 37 years of age with their autologous SVBT cryopreserved on day 5 (5d-SVBT, n = 737) or day 6 (6d-SVBT, n = 154) and DVBT on day 5 (5d-DVBT, n = 129) or day 6 (6d-DVBT, n = 31) from January 2009 to December 2011.

Results

The clinical pregnancy rate (41.8 % vs. 48.1 %, p = 0.184) and ongoing pregnancy rate (36.6 % vs. 45.0 %, p = 0.072) were not significantly different between the 5d-SVBT group and the 5d-DVBT group. However, the clinical pregnancy (29.9 % vs. 58.1 %, p = 0.003) and ongoing pregnancy rates (23.4 % vs. 51.6 %, p = 0.001) were significantly lower in the 6d-SVBT group compared with those in the 6d-DVBT group. The implantation rate (42.2 % vs. 34.5 %, p = 0.03) of the 5d-SVBT group was significantly higher than that of the 5d-DVBT group, while the implantation rate (29.9 % vs. 37.1 %, p = 0.303) of the 6d-SVBT group was not statistically different compared with that in the 6d-DVBT group. The multiple pregnancy rates (1.0 % in the 5d-SVBT group vs. 38.7 % in the 5d-DVBT group, p < 0.001 and 0 % in the 6d-SVBT group vs. 22.2 % in the 6d-DVBT group, p = 0.001) were statistically significantly lower in the SVBT group compared with those in the DVBT group regardless of the day of vitrification.

Conclusions

This study showed that the 5d-SVBT resulted in comparable clinical outcomes compared to the 5d-DVBT while the 6d-SVBT yielded significantly lower clinical outcomes compared to the 6d-DVBT.     

Frozen blastocyst transfer reduces incidence of ectopic pregnancy compared with fresh blastocyst transfer: a meta-analysis

Background: Ectopic pregnancy (EP) is the main cause of maternal death during early pregnancy. Blastocyst transfer (BT) reduces EP rates compared with cleavage stage embryo transfer (ET), and frozen ET reduces EP rates compared with fresh ET. However, data comparing the EP rate of fresh BT and frozen BT are limited. The objective of this meta-analysis was to determine whether frozen BT decreases the EP rate compared with fresh BT.

Materials and methods: PUBMED, EMBASE, and Cochrane Library databases were searched for papers with no limitation on language and publication year. A systemic literature search identified 14 studies which met the inclusion criteria for further analysis. EP rate is our only outcome measure. All statistical analyses were carried out using RevMan software [version 5.3]. Random effects models were chosen to calculate the odds ratio (OR).

Results: Fourteen retrospective studies (n?=?251,762 cycles) were finally analyzed, including 154,214 and 97,548 cycles undergoing fresh and frozen BT, respectively. Compared with fresh BT, the frozen BT was associated with an decrease in EP rate [OR?=?1.79; 95% confidence interval (CI)?=?1.37–2.34, heterogeneity was 71%], the EP rate was 1.74% (2683/154,214 pregnancies) for fresh BT and 0.97% (949/97,548 pregnancies) for frozen BT. The number of embryos transferred in subgroup analysis found that the incidence of EP in frozen BT is lower than fresh BT (OR?=?1.62; 95% CI?=?1.38–1.91), the EP rate was 1.2% (206/16,610 pregnancies) for one fresh BT and 0.8% (153/19,569 pregnancies) for one frozen BT, also the EP rate was 1.9% (78/4043 pregnancies) for two fresh BT and 1.1% (19/1790 pregnancies) for two frozen BT.

Conclusions: Our meta-analysis found that frozen BT was associated with an decrease in EP rate compared with fresh BT in IVF/ICSI patients. We also found that a single frozen BT is the better choice as a method to reduce the incidence of EP.     

Embryo fragmentation as a determinant of blastocyst development in vitro and pregnancy outcomes following embryo transfer

OBJECTIVE(S): To determine how the type of embryo fragmentation on day 3 affects progression of human embryos to blastocyst and pregnancy rates following embryo transfer. STUDY DESIGN: Retrospective analysis of all in vitro fertilization cycles in patients < or =40 years of age or younger from January 2002 through December 2003, during which time surplus day 3 embryos were transferred to blastocyst medium for extended culture. All embryos (4 cells or more) not suitable for transfer or freezing 72 hours following in vitro fertilization were placed into microdroplets (60 microL) of blastocyst medium and cultured for an additional 48 hours to assess blastocyst formation. Normal blastocyst development required blastulation, a visible inner-cell mass, trophectoderm cells covering 60% of the inner zona surface and thinning of the zona. The rate of blastocyst formation was then analyzed (chi 2 and analysis of variance) against the type of fragmentation 72 hours after insemination. Pregnancy outcomes were analyzed with respect to the pattern of fragmentation in cleaving embryos transferred after 3 days of culture. RESULTS: A total of 1566 embryos were cultured beyond day 3 of development of which 229 (14.6%) reached the blastocyst stage and were frozen. Embryos exhibiting no fragmentation or type I fragmentation had significantly higher blastocyst development rates (27.9% and 19.9%) than embryos with type 2 or 3 fragmentation (13.9 and 8.8, respectively; P < .001). No embryos with type 4 or 5 fragmentation progressed to blastocyst. The average type of fragmentation in transferred embryos correlated with pregnancy outcome and embryo age. CONCLUSION(S): More pervasive embryo fragmentation was associated with a decreasing rate of blastocyst development with day 3 embryos. To the extent that blastocyst development rates of day 3 embryos is an index of embryo viability, our findings establish that careful classification of the type of embryo fragmentation is important in selection of day 3 embryos for transfer. Recent reports of associations among embryo fragmentation, aneuploidy, apoptosis, and patient age support these conclusions.     

Retrospective clinical analysis of two artificial shrinkage methods applied prior to blastocyst vitrification on the outcome of frozen embryo transfer

Purpose

Vitrification significantly improves the rates of blastocyst survival and clinical pregnancy following frozen embryo transfer (FET). However, ice crystal formation during the freezing process reduces the blastocyst survival rate. Artificial shrinkage (AS) prior to blastocyst vitrification decreases the formation of ice crystals, increasing the blastocyst survival rate. The aim of this study was to identify an efficient AS method to improve blastocyst survival rates following vitrification.

Method

Use of the 29-gauge needle AS and Laser pulse AS methods prior to vitrification was compared in terms of the impacts on the rates of blastocyst survival in FET cycles, blastocyst hatching, clinical pregnancy after transfer, embryo implantation, abortion, gestational duration and birth weight.

Result

In total, 438 blastocysts in 219 cycles were thawed, resulting in survival of 407 (92.9 %). Of these, 213 cycles were transferred, resulting in 129 clinical pregnancies (60.6 %) and 140 successful births. There were no differences between the two methods in the rates of blastocyst survival, clinical pregnancy, embryo implantation and abortion. However, the 29-gauge needle AS group was associated with a significantly lower blastocyst hatching rate (83.6 % vs. 91.2 %), shorter average gestational duration (37.36 ± 2.34 vs. 38.06 ± 1.76), and higher premature birth rate (40.00 % vs. 21.15 %) compared with Laser pulse AS group.

Conclusion

No significant differences in the effectiveness of the two methods applied prior to blastocyst vitrification were observed before birth, while after birth, a significantly improved clinical outcome was obtained with laser pulse AS indicating that this is a more effective pre-processing method for blastocyst vitrification.     

Comparison of the sex ratio with blastocyst transfer and cleavage stage transfer

Purpose : To evaluate the sex ratio in births conceived with blastocyst transfer compared to day 3-ET. Methods : A retrospective analysis of IVF patients who became pregnant after blastocyst or cleavage stage transfer at Stanford University Hospital and a literature review were performed. Result(s) : In the day 3-ET group, the male-to-female (M/F) ratio was 157/139 (53%/47%) compared to 97/66 (59.5%/40.5%) in the blastocyst group (P = 0.18). Similar trends have been found in individual studies in the literature but reached statistical significance in only one out of six reports reviewed. The combined data from our study and the literature show a male-to-female ratio of 797/594 (57.3%/42.7%) in blastocyst transfer compared to 977/932 (51.2%/48.8%) in day 3-ET (P = 0.001). Conclusion(s) : Although individual studies may lack power to show an altered sex ratio with blastocyst transfer, the combined data presented in this report do suggest that the M/F ratio is higher with blastocyst transfer compared to cleavage stage transfer.     

Clinical and pregnancy outcomes of double and single blastocyst transfers related with morphological grades in vitrified-warmed embryo transfer

ObjectiveTo evaluate clinical and pregnancy outcomes of double and single blastocyst transfers related with morphological grades in vitrified-warmed embryo transfer.Materials and methodsIn a retrospective cohort analysis, data were assessed from women who underwent vitrified-warmed blastocyst transfers (VBT) at CHA Gangnam Medical Center between 2014 and 2015. All VBT cycles were categorized into three groups according to blastocyst quality: GG (double good blastocysts transfer), GP (one good and one poor blastocyst transfer), and GS (single good blastocyst transfer). Blastocysts were graded morphologically and ⩾3BB grade was considered good quality.ResultsThere were 628 transfers in group GG, 401 transfers in group GP, and 277 transfers in group GS. Both clinical pregnancy rate (CPR) and live birth rate (LBR) were the highest in group GG (CPR 65.9%, LBR 55.3%, p < 0.001), but not significantly different between group GP and GS. Multiple pregnancy rates increased significantly in the following order: GS (1.4%), GP (13.5%), and GG (25.6%). Single LBR was the highest in group GS (38.6%, p < 0.001).ConclusionAs an effective VBT, especially for reducing multiple pregnancy and increasing single live birth, single good blastocyst transfer may be recommended rather than any double blastocyst transfer methods. Moreover, transferring a good and a poor blastocyst simultaneously should be avoided.     

Post-warming embryo morphology is associated with live birth: a cohort study of single vitrified-warmed blastocyst transfer cycles

PurposeThis study aims to examine whether blastocyst morphology post-warming correlates with live birth.MethodsIn this cohort study, morphological characteristics post-warming were reviewed in all single vitrified-warmed blastocyst transfer cycles performed between November 2016 and May 2017. Immediately before transfer, the degree of blastocoel re-expansion was graded as A, fully expanded; B, partially expanded ≥ 50%; C, partially expanded < 50%; and D, collapsed. The degree of post-warming cell survival was graded on a scale of 50 to 100% and was then classified into 4 groups: very low 50–70%, low 71–80%, moderate 81–90%, and high 91–100%.ResultsOverall, 612 cycles were reviewed, of which 196 included PGT-A tested embryos. The live birth rate (LBR) increased from 11.4% in the collapsed blastocysts group to 38.9% in the post-warming full re-expansion group (p < 0.001) and from 6.5% for blastocysts with a very low cell survival rate to 34.7% for blastocysts with high cell survival rate (p = 0.001). LBR was 6.7% for blastocysts with the worst post-warming morphological characteristics, namely, collapsed with very low cell survival rate. On multivariate analyses, partial blastocyst re-expansion ≥ 50%, full re-expansion, and high cell survival rate remained significantly associated with live birth, after controlling for female age, pre-vitrification morphological grading, and PGT-A. A sub-analysis of cycles using PGT-A tested embryos showed similar results.ConclusionPost-warming re-expansion and high cell survival rate are associated with higher LBR in euploid and untested blastocysts. However, embryos with poor post-warming morphology still demonstrate a considerable probability of live birth, and they should not be discarded.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10815-021-02390-z.     

A quantitative approach to blastocyst quality evaluation: morphometric analysis and related IVF outcomes

Purpose

To quantify blastocyst morphologic parameters with a feasible and standardized tool, investigating their predictive value on implantation outcome.

Method

The study retrospectively analyzes 124 blastocysts from 75 patients. Quantitative measurements of blastocyst expansion, inner cell mass and trophoectoderm were taken using digital image analysis software.

Result(s)

Blastocysts areas were found to be ranging from 11626.2 up to 35076.4 μm². The area of an early blastocyst is A ≤ 18500 μm² with a mean diameter d = 140 ± 9 μm, and the area of an expanded blastocyst is A ≥ 24000 with d = 190 ± 9 μm. While blastocyst mean area was not related to implantation rate, more expanded blastocysts displayed a significantly higher implantation rate. Trophoectoderm cell number is a predictor of positive outcome: since a higher of cells (25.6 ± 11.3 vs 16.3 ± 12.8) `forming a tightly knit epithelium is prognostic of implantation potential. Conversely, inner cell mass size is significantly related to implantation only in expanded blastocysts (3122.7 ± 739.0 vs. 2978.1 ± 366.0 μm²).

Conclusion(s)

Evaluation of blastocyst morphology with a digital image system could be a valuable tool to standardize blastocyst grading based on quantitative parameters. Therefore, digital analysis may be helpful in identifying the best blastocyst to transfer.     

Comparison of the pregnancy and obstetric outcomes between single cleavage-stage embryo transfer and single blastocyst transfer by time-lapse selection of embryos

To compare the pregnancy and obstetric outcomes following single cleavage-stage embryo transfer (SCT) and single blastocyst transfer (SBT) using time-lapse imaging (TLI), a total of 2066 normally fertilized and cleaved embryos from 233 patients were divided into Day 3 SCT group (n?=?171) and Day 5 SBT group (n?=?62) according to patient’s willingness. Embryo selection criteria were based on embryo cleavage patterns, timing parameters, and blastocyst quality. The pregnancy and obstetric outcomes of each group were evaluated. There were no statistically significant differences with regard to pregnancy outcomes including the implantation rate, early abortion rate, ongoing pregnancy rate and live birth rate, and obstetric outcomes including preterm birth rate, gestational week, birth height, birth weight and fetal malformation rate between SCT group and SBT group. SBT group had significantly higher monozygotic twinning (MZT) rates than SCT group (6.98% vs. 0, p?<?.05). Although not statistically significant, there was a trend of higher proportion of male-to-female sex ratio at birth in SBT group than SCT group (1.38 vs. 1.05). Based on the combination of cleavage patterns and timing parameters, SCT may be an alternative to SBT because it can provide similar pregnancy and obstetric outcomes and meanwhile lower monozygotic twinning rates.     

Case report: successful delivery following the transfer of a human re-vitrified day-7 spontaneously hatched blastocyst developed from vitrified cleaved embryos

Purpose  

To report a successful delivery after the transfer of a re-cryopreserved day-7 hatched blastocyst.     

A successful birth of healthy monozygotic dichorionic diamniotic (DD) twins of the same gender following a single vitrified-warmed blastocyst transfer

Purpose  

To report the world’s first case of live monozygotic dichorionic diamniotic (DD) twins after a single vitrified-warmed blastocyst transfer in a cycle during which hormone supplements were given.     

新鲜周期单囊胚移植314例临床结局

目的 探讨新鲜周期单囊胚移植(single blastocyst transfer,SBT)的可行性。方法 回顾性分析314个新鲜周期行SBT患者的临床资料,从年龄、囊胚分级等方面比较分析临床结局。结果 妊娠组患者的年龄显著低于未妊娠组(P0.001);选择性SBT(elective SBT,eSBT)组的临床妊娠率显著高于非选择性SBT(non-elective SBT,non-eSBT)组(P0.05);年龄≤35岁的患者,eSBT组的临床妊娠率与双囊胚移植(double blastocyst transfer,DBT)组无统计学差异,但多胎妊娠率显著降低(P0.001);而35岁的患者,eSBT组的临床妊娠率低于DBT组(P0.05),组间的多胎妊娠率无统计学差异(P0.05);囊胚扩张和孵化的程度与临床妊娠率显著相关,优质囊胚可获得更好的妊娠结局。结论 妊娠结局与囊胚的质量及患者的年龄显著相关,对于年龄≤35岁,有优质囊胚的患者,可以进行SBT。     

Molecular and cellular events involved in the completion of blastocyst implantation

Blastocyst implantation is an interactive process between the embryo and the uterus. The synchronization of embryonic development with uterine differentiation to a receptive state is essential for a successful pregnancy. The period of uterine receptivity for implantation is limited. Although implantation involves the interaction of numerous signaling molecules, our understanding of the hierarchical mechanisms that coordinate with the embryo–uterine dialogue is not yet sufficient to prevent infertility caused by implantation failure. This review highlights our knowledge on uterine receptivity and hormonal regulation of blastocyst implantation in mice. We also discuss the adhesion molecules, cross-linker proteins, extracellular proteins, and matricellular proteins involved in blastocyst implantation. Furthermore, our recent study reveals that selective proteolysis in an activated blastocyst is associated with the completion of blastocyst implantation after embryo transfer. A better understanding of uterine and blastocyst biology during the peri-implantation period would facilitate further development of reproductive technology.     

Blastocyst age,expansion, trophectoderm morphology,and number cryopreserved are variables predicting clinical implantation in single blastocyst frozen embryo transfers in freeze-only-IVF

PurposeTo determine which blastocyst assessment variables predict clinical implantations in single blastocyst frozen embryo transfers (FET) of freeze-only-IVF cycles, following improved vitrified-warmed blastocyst survival and developmental competence preservation.MethodIn this retrospective cohort study performed at a single private IVF center, the pregnancy outcomes of 1795 single blastocyst FET cycles were analyzed, from freeze-only-IVF retrievals performed between January 2017 and January 2020. Stepwise forward logistic regressions with clinical implantation (i.e., normal gestational sac and cardiac activity) as dependent variable were performed to identify the significant predictors. All blastocysts were vitrified using Cryotop technology, with before transfer (post-warming) blastocyst morphology scores used in all analyses.Result(s)The 1795 blastocysts transferred were vitrifıed on embryo days 4 (1057), 5 (716), and 6 (22). The overall clinical implantation rate was 50.9%; however, using blastocyst age and blastocyst morphological score the clinical implantation rates increased from 49.0% (day-4 1 and 2) and 25.2% (day-5 1 and 2) to 71.2% (day-4 4AA) and 64.3% (day-5 4AA), respectively. Whereas full (≥3) blastocysts with scores of AA and BA had similar clinical implantation rates (66.2 vs. 66.7%), the rate of full blastocysts with scores of AB was lower (58.9%). In stepwise forward logistic regressions, female age, blastocyst age, blastocyst expansion score, blastocyst trophectoderm score, and number of blastocysts vitrified were significant predictors of clinical implantation.Conclusion(s)Using blastocyst age and before transfer blastocyst expansion and trophectoderm morphology scores to select blastocysts, clinical implantation rates greater than 70% could be achieved for top-scoring blastocysts.     

Comparison of blastocyst transfer to day 3 transfer with assisted hatching in the older patient

OBJECTIVE: To compare cycle outcomes in similar populations of women over 40 who underwent blastocyst transfer compared with women who had day 3 embryo transfer with assisted hatching (ET/AH). DESIGN: Retrospective study. STTING: University hospital-based program. PATIENT(S): Eighty-six IVF cycles in women ages 40 to 43 years who had more than three eight-cell embryos on day 3.On day 3 of embryo culture, patients chose either to undergo blastocyst transfer or day 3 ET/AH. MAIN OUTCOME MEASURE(S): Pregnancy and cryopreservation rates were recorded. RESULT(S): In 48 cycles, blastocyst transfer was performed, and in 38 cycles day 3 ET/AH was performed. There was no statistically significant difference between the blastocyst transfer group and the day 3 ET/AH group with respect to age (41.1 +/- 0.9 years vs. 41.6 +/- 0.8 years), percentage of intracytoplasmic sperm injection cycles (29.2% vs. and 27.6%), number of oocytes (14.9 +/- 5.6 vs. 12.8 +/- 4.0), or number of eight-cell embryos (6.1 +/- 2.2 vs. 5.4 +/- 1.5). Significantly fewers embryos were transferred per cycle with blastocyst transfer (2.6 +/- 1.0) compared with day 3 ET/AH (5.9 +/- 2.0). The viable pregnancy rate was similar in the blastocyst transfer group (29.2%) and in the day 3 ET/AH group (26.3%). Embryos for cryopreservation were available in significantly more cycles in the blastocyst transfer group (52.1%) than in the day 3 ET/AH group (21.1%). Cleavage stage arrest occurred only in one cycle. CONCLUSIONS: Blastocyst transfer appears to be as effective as day 3 ET/AH in older patients with good embryos. Higher cryopreservation rate in the blastocyst transfer group may represent an advantage over day 3 ET/AH. Older women may also benefit from the information that extended culture provides them regarding their oocyte quality.