P–259 Blastocyst quality is associated with both the start and the duration of compaction after ICSI

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
K Wouters ◽  
L Va. Landuyt ◽  
M Regin ◽  
H Tournaye ◽  
G Verheyen ◽  
...  
Keyword(s):  
Embryo Quality ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Total Duration ◽  
Time Lapse ◽  
Poor Quality ◽  
Blastocyst Stage ◽  
Human Embryos ◽  
The Poor ◽  
Blastocyst Quality

Abstract Study question Is the start and the total duration of compaction related to embryo quality? Summary answer The timing of the start, the end and the total duration of compaction are associated with blastocyst quality grade in the IVF laboratory. What is known already Preimplantation embryo development follows a programmed timeline during which a series of critical events take place. One event typically occurring on day 3/4 post fertilisation is the formation of adherence junctions between blastomeres in a process called compaction. It is considered the first morphological event in the differentiation process of the mammalian embryo. Evaluation of developmental events are used to optimize the selection of the most competent embryos for transfer and/or cryopreservation in the IVF laboratory. It has already been shown that the time of full compaction is indicative for high-quality blastocysts with a higher implantation rate. Study design, size, duration A single-centre retrospective observational study including 74 ICSI cycles performed in 2020. Injected oocytes were cultured in blastocyst medium (Origio) in the EmbryoScope + (Vitrolife) for 5/6 days. Embryos that reached the blastocyst stage were evaluated for the start of compaction, the time to reach full compaction and the total duration of compaction. These parameters were compared between good- and poor-quality blastocysts; the primary outcome parameter of the study was embryo quality. Participants/materials, setting, methods Only ICSI cycles with ejaculated fresh/frozen-thawed sperm and monitored in time-lapse incubator were included. All MNC, IVM and PGT cycles were excluded. Time zero was the start of ICSI. Good-quality embryos were full and expanded blastocysts with good-quality inner cell mass and trophectoderm (AA, AB, BA and BB according to Gardner and Schoolcraft (1999)). GraphPad Prism was used for statistical analysis. After testing for normality and homogeneity, unpaired t-test or Mann-Whitney test determined significant differences. Main results and the role of chance In this study, of the 528 included 2PN oocytes, 229 (43.4%) reached the blastocyst stage and 299 (56.6%) were arrested. Among the former, 131 (57.2%) blastocysts were classified in the good-quality group and 98 (42.8%) blastocysts in the poor-quality group. In general, human embryos compacted slowly while dividing further and the blastomeres moved during the compaction process. The start of compaction was heterogeneous (between 50.9 and 102.7 hours post ICSI; mean=80.0 hours), as well as the cell number at the initiation (between 4 and 18 blastomeres; mean=12 blastomeres). The time analysis showed that the embryos in the good-quality group started to compact significantly earlier than those in the poor-quality group (mean=78.6 vs 82.2 hours; R²=0.06; p < 0.01). We confirmed that blastocysts in the good-quality group reached full compaction earlier than those in the poor-quality group (mean=86.8 vs 93.8 hours; R²=0.17; p < 0.01). Furthermore, the total duration of compaction was significantly lower in the good-quality than in the poor-quality group (median=7.4 vs 10.7 hours; p < 0.01). Limitations, reasons for caution As this is a retrospective study, the influence of uncontrolled variables cannot be excluded. The absence of the pregnancy outcome and live birth rate is a shortcoming and will be subject of a larger patient-to-patient study. Wider implications of the findings: These results indicate that an earlier start and a shorter duration of compaction are associated with better blastocyst quality. These morphological events can be valuable additional parameters in selecting the embryo of better quality when using a time-lapse incubator. Trial registration number Not applicable

scholarly journals Transfer of hatched blastocyst can result in skewed sex ratio: a retrospective cohort study

2021 ◽  
Author(s):  
Jinliang Zhu ◽  
Ying Lian ◽  
Xinjie Zhuang ◽  
Shengli Lin ◽  
Xiaoying Zheng ◽  
...  
Keyword(s):  
Sex Ratio ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Poor Quality ◽  
Blastocyst Stage ◽  
Neonatal Outcomes ◽  
Blastocyst Transfer ◽  
Female Ratio ◽  
Single Blastocyst Transfer ◽  
Blastocyst Quality

Abstract Background The correlation between blastocyst quality and birthweight, neonatal outcomes is still controversial. There is a significantly higher male: female ratio among good quality blastocysts (advanced trophoderm morphology) but in the expansion degree, the significance for sex ratio is unclear. Methods A total of 617 and 6803 live singleton births resulting from the transfer of fresh and frozen-thawed single blastocysts in the Reproductive Medicine Center of Peking University Third Hospital from 2009 to 2020 were included. Live singleton births from fresh and frozen-thawed single blastocyst transfer were stratified by inner cell mass/trophoderm morphology and degree of blastocoel expansion. Multivariate linear regression was used to analyze the correlation between expansion, inner cell mass/trophoderm morphology, and birthweight, Z score, gestational weeks. Logistic regression was used to analyze the relationship between expansion, ICM/TE morphology and sex, neonatal outcomes. Results There was no significant correlation between birthweight, neonatal outcomes and blastocyst quality in fresh and frozen-thawed single blastocyst transfer cycles. However, the proportion of male infants in the hatched blastocyst (stage-6) group (67.9% vs. 54.2%; p < 0.001) [OR: 1.76 95% CI (1.34–2.32)] and hatching blastocyst (stage-5) group (61.7% vs. 54.2%; p = 0.001) [OR: 1.36 95 C.I (1.14ཞ1.62)] was significantly higher than that in the expanded blastocyst (stage-4) group. Conclusions The transfer of poor-quality blastocysts is unlikely to affect birthweight and neonatal health; however, transfer of stage-6 blastocysts can result in extremely skewed sex ratio.

P–132 Centralized versus local embryologists scoring of blastocyst quality obtained in a large European multicenter clinical trial

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Montag ◽  
E Va. de. Abbeel ◽  
T Ebner ◽  
P Larsson ◽  
B Mannaerts
Keyword(s):  
Quality Assessment ◽  
Data Storage ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Time Lapse ◽  
Ongoing Pregnancy Rate ◽  
Quality Classification ◽  
Blastocyst Quality ◽  
Selection Of ◽  
Local Assessment

Abstract Study question Does blastocyst quality scoring by central assessment deviate from local assessment and potentially lead to the selection of a different single blastocyst for transfer? Summary answer Central and local assessment provided the same quality classification (poor / good / top) in 69% of all blastocysts and 63% of all transferred blastocysts. What is known already Blastocyst quality is scored most frequently by three morphological parameters, namely expansion and hatching (EH) status, inner cell mass (ICM) grading and trophectoderm (TE) grading. The score is used to define the quality classification (poor / good / top) which determines which embryo is to be transferred or cryopreserved. Blastocyst scoring and grading can be highly subjective, which does influence the choice for transfer and cryopreservation. Time-lapse imaging technology captures additional input about embryo development as well as enables centralized data storage and sharing for independent central assessments. Study design, size, duration Pooled embryo analysis from a prospective, randomized, multicenter trial (RAINBOW) of 619 women undergoing ovarian stimulation with an individualized dose of follitropin delta in a long GnRH agonist protocol between May 2018 and January 2020. Blastocysts were centrally assessed using time-lapse images by two independent assessors and one adjudicator . Selection of the blastocyst for transfer by local assessment was based on morphological scoring and not on morphokinetic time-lapse parameters. Participants/materials, setting, methods Oocytes were fertilized by ICSI and cultured in the Embryoscopeâ (Vitrolife) up to day 5 for transfer or day 5/6 for cryopreservation. Embryos were assessed as either non-blastocyst or blastocyst. Blastocysts were graded centrally and locally at 116 hrs of development, based on EH status (1–6), ICM (A-D) and TE grading (A-D). Central assessors were blinded to local assessment and embryo transfer selection. Main results and the role of chance In total 4282 embryos were assessed centrally, of which 2046 day 5 embryos (48%) were adjudicated due to a scoring difference of at least one parameter between the two central assessors. In total 38% of day 5 embryos were judged as non-blastocysts and 62% as blastocysts of which 61% (i.e. 38% of all embryos) were determined to be of good or top quality. Identical results in terms of quality classification (poor / good / top) were obtained for 69% of blastocysts between local and central assessment and in 78%, between the two central assessors. Moreover, central and local scoring were identical in 62% for EH status, 53% for ICM grading and 57% for TE grading. For all transferred blastocysts (n = 508), central and local quality assessment was aligned for 63%. The ongoing pregnancy rate following single blastocyst transfer (SBT) was 41% (202/489), and similar to when considering only the transfers for which the central assessment had the same or a higher classification than the local assessment (166/411=40%). In 16% of all SBT, central quality assessment gave a lower score for the transferred blastocyst than the central assessment. This discrepancy could potentially have led to transfer of a different blastocyst. Limitations, reasons for caution This trial included assessments made by embryologists from 20 IVF centres. Some centres has limited experience with time-lapse technology for morphological blastocyst scoring. Scoring could therefore have been affected by differences in focal planes, magnification and contrast compared to inverted microscopy, with potential influence on blastocyst scores and quality classification. Wider implications of the findings: Local and central blastocyst quality classification based on morphology aligns well but remains subjective. Embryo assessment may benefit from using tools like artificial intelligence-based algorithms for a more objective analysis. Trial registration number NCT03564509

scholarly journals Does Blastocyst Morphology Influence Live Birth Rate after Frozen-thawed Single Euploid Embryo Transfer?

2021 ◽  
Author(s):  
Na Li ◽  
Yichun Guan ◽  
Bingnan Ren ◽  
Yuchao Zhang ◽  
Yulin Du ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Live Birth ◽  
Birth Rate ◽  
Inner Cell Mass ◽  
Medical Center ◽  
Cohort Analysis ◽  
Cell Mass ◽  
Live Birth Rate ◽  
Poor Quality ◽  
Blastocyst Quality

Abstract Objective To investigate whether the morphologic parameters of euploid blastocyst influence the live birth rate (LBR) following single frozen-thawed embryo transfer (FET) cycles? Methods A retrospective cohort analysis involving autologous single FET cycles after preimplantation genetic testing for aneuploidy (PGT-A) through next generation sequencing (NGS) by a university-based reproductive medical center that was performed from June 2017 to September 2019.Women were divided into three age groups (< 30, 30–34 and ≥ 35 years). The primary outcome measure was LBR. Outcomes were compared to determine the association between different blastocyst quality (Good, Average and Poor), inner cell mass (ICM) grade (A and B), and trophectoderm (TE) grade (A, B and C) and LBR. Results We included 232 single FET cycles for analysis, the total LBR was 48.48%. In the youngest group (< 30 years, n = 86), LBR were compared between cycles with various blastocyst quality (72.22% for good quality, 54.55% for average quality and 34.78% for poor quality; P = 0.019), ICM grade (70.59% for grade A and 42.03% for grade B; P = 0.035) and TE grade (85.71% for grade A,57.58% for grade B and 34.78 for grade C; P = 0.015). Similarly, in the 30–34 years group, LBR ranged from 50.00% for good quality to 45.45% for poor quality (P = 0.870), from 35.29% for ICM grade A to 51.22% for ICM grade B (P = 0.291), from 60.00% for TE grade A to 45.45% for TE grade C (P = 0.634). Likewise, in the oldest group (≥ 35years, n = 47), LBR were also comparable between these subgroups, but no significant differences were seen in blastocyst morphologic parameters and LBR (P > 0.05). Conclusion After single FET cycles, the LBR was associated with morphologic parameters of euploid blastocysts, especially in women < 30 years old. However, these differences were not found in women older than 30 years. We suggested that for older women whose embryos undergoing PGT-A with NGS to be euploid have the same development potential regardless of their blastocyst morphology.

The human blastocyst: cell number, death and allocation during late preimplantation development in vitro

Development ◽  
1989 ◽  
Vol 107 (3) ◽  
pp. 597-604 ◽  
Author(s):  
K. Hardy ◽  
A.H. Handyside ◽  
R.M. Winston
Keyword(s):  
Cell Death ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Cell Number ◽  
Blastocyst Stage ◽  
Human Embryos ◽  
Cell Numbers ◽  
Human Blastocyst ◽  
Development In Vitro

The development of 181 surplus human embryos, including both normally and abnormally fertilized, was observed from day 2 to day 5, 6 or 7 in vitro. 63/149 (42%) normally fertilized embryos reached the blastocyst stage on day 5 or 6. Total, trophectoderm (TE) and inner cell mass (ICM) cell numbers were analyzed by differential labelling of the nuclei with polynucleotide-specific fluorochromes. The TE nuclei were labelled with one fluorochrome during immunosurgical lysis, before fixing the embryo and labelling both sets of nuclei with a second fluorochrome (Handyside and Hunter, 1984, 1986). Newly expanded normally fertilized blastocysts on day 5 had a total of 58.3 +/− 8.1 cells, which increased to 84.4 +/− 5.7 and 125.5 +/− 19 on days 6 and 7, respectively. The numbers of TE cells were similar on days 5 and 6 (37.9 +/− 6.0 and 40.3 +/− 5.0, respectively) and then doubled on day 7 (80.6 +/− 15.2). In contrast, ICM cell numbers doubled between days 5 and 6 (20.4 +/− 4.0 and 41.9 +/− 5.0, respectively) and remained virtually unchanged on day 7 (45.6 +/− 10.2). There was widespread cell death in both the TE and ICM as evidenced by fragmenting nuclei, which increased substantially by day 7. These results are compared with the numbers of cells in morphologically abnormal blastocysts and blastocysts derived from abnormally fertilized embryos. The nuclei of arrested embryos were also examined. The number of TE and ICM cells allocated in normally fertilized blastocysts appears to be similar to the numbers allocated in the mouse. Unlike the mouse, however, the proportion of ICM cells remains higher, despite cell death in both lineages.

104 BIRTH OF FIRST BUFFALO (BUBALUS BUBALS) CALF FOLLOWING EMBRYO SPLITTING AND POLYMERASE CHAIN REACTION SEXING AT BLASTOCYST STAGE

2012 ◽  
Vol 24 (1) ◽  
pp. 164 ◽  
Author(s):  
M. Zhang ◽  
H. H. Chen ◽  
J. W. Tang ◽  
X. W. Liang ◽  
M. T. Chen ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Blastocyst Stage ◽  
Buffalo Calf ◽  
Practical Procedure ◽  
Blastocyst Quality ◽  
Grade 3 ◽  
Vitro Development

Embryo-splitting technology provides an effective procedure for increasing the number of transferable embryos per donor, producing genetically identical offspring and facilitating embryo sexing. The ability to identify the sex of embryos before transfer will offer a reliable, economical and practical procedure for buffalo breeding. In this study, we have assessed the feasibility of production of offspring with controlled sex in buffalo by first comparing the effect of blastocyst quality on the viability of demi-embryos and then identifying the sex of a demi-embryo by multiplex-nested PCR before transfer into the recipient. In vitro-matured buffalo oocytes were fertilized by IVF and cultured to the blastocyst stage for 6 to 7 days as described by Lu et al. (2007 Anim. Reprod. Sci. 100, 192–196). These blastocysts were classified in terms of their developmental pattern and morphology on a scale of 1 to 3 grades as described by McEvoy et al. (1990 Theriogenology 33, 1245–1253). Blastocysts were split into 2 equal parts by a micromanipulation system. Viability of the resulting demi-embryos was confirmed by formation of a blastocoel cavity and definite inner cell mass after culture for 24 h. One of the zone-free demi-embryos derived from a grade-1 blastocyst was cultured in TCM 199 supplemented with 10% fetal bovine serum for another 2 h, then was transplanted to a spontaneous oestrous recipient. The other demi-embryo was used for sexing by multiplex-nested PCR (Fu et al. 2007 Theriogenology 68, 1211–1218). The results showed that grade-1 blastocysts yielded more viable demi-embryos than grade-2 and grade-3 blastocysts [P < 0.01; 73/92 (79.67%) vs 32/76 (47.05%) vs 26/94 (26.53%), respectively]. Transplantation of the presumed-Y demi-embryo derived from grade-1 blastocyst into a recipient resulted in the birth of a male buffalo calf. To the best of our knowledge, this is the first buffalo calf produced following embryo splitting and PCR sexing of the embryo at the blastocyst stage. Successful birth of the desired-sex offspring in the present study indicates the feasibility of using embryo splitting in combination with multiplex-nested PCR sexing to produce offspring of controlled sex in swamp buffalo. However, the quality of embryos before splitting was an important factor governing the in vitro development of viable demi-embryos. This study was supported by the Guangxi Science and Technology R&D Program (0626001-3-1, 0815008-2-4).

155 CHARACTERIZATION OF LIPID DROPLETS IN BOS INDICUS AND BOS TAURUS EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 226 ◽  
Author(s):  
E. P. López-Damián ◽  
T. Fiordelisio ◽  
M. A. Lammoglia ◽  
M. Alarcón ◽  
M. Asprón ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Embryo Quality ◽  
Developmental Stages ◽  
Bos Taurus ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Bos Indicus ◽  
Nile Red ◽  
Blastocyst Stage ◽  
Transfer Program

Accurate evaluation of bovine embryos for assessing developmental stage and quality is critical to the success of any embryo transfer program. However, this evaluation process has been reported to be highly subjective in Bos indicus (BI) and can vary as much as 23% compared with that of Bos taurus (BT). These differences in assessment may be related to the quantity of lipid droplets (LD) within the embryo, which has been shown to have a negative effect in cryopreserving embryos. The aim of the present study was to characterize the number and size of LD in different developmental stages of fresh embryos from BI and BT and to compare LD across the three different embryo quality grades (1 = excellent or good, 2 = fair, and 3 = poor). Nonsurgical embryo collection was performed 7 days post-insemination in 10 BI and 10 BT females. Forty-eight embryos were evaluated for stage and grade using stereoscopic microscopy, processed for transmission electron microscopy, and stained with Nile red. Digitalized images were analyzed with ImageJ (National Institutes of Health, Bethesda, MD, USA), contour of lipid droplets were designed, and values of perimeter, area, and fluorescence intensity were assessed. Nonparametric statistical analysis (Mann–Whitney) was utilized. There was no difference in LD number for BT or BI for morulae and blastocyst; however, BI morulae presented larger LD compared with blastocyst stage embryos (286 µm2 v. 223 µm2; P < 0.05). Likewise, BI TF cells had more LD compared with inner cell mass (ICM) cells (48 v. 36; P < 0.05). BT TF cells exhibited larger LD compared with ICM cells (149 µm2 v. 128 µm2; P < 0.05), while BI embryos exhibited a larger area of LD in the ICM compared with the TF (591 µm2 v. 472 µm2; P < 0.05). In all embryos, BI contained more lipid droplets than BT (78 v. 49; P < 0.05). Across all quality grades (good, fair, and poor) there was no difference in the number of LD in BT embryos; however, BI grade-3 embryos presented more LD than grade-1 (36 v. 25). BT embryos LD were larger than BI LD (907 µm2 v. 625 µm2; P < 0.05). Fluorescence images showed higher arbitrary units of fluorescence (auf) for LD in BI. Compared with BT embryos (386 auf v. 280 auf; P < 0.05). These results suggest that BI embryos contain more and smaller LD than BT embryos and the LD described for BI embryo quality grade 1 are larger than those of quality grades 2 and 3, and even though the number of LD in morulae and blastocyst stage embryos are not different LD size is reduced as development occurs. Research funding provided by UNAM-DGAPA-PAPIIT IN200810.

6 ANEUPLOIDY TOLERANCE IN RHESUS MACAQUE PRE-IMPLANTATION EMBRYOS VIA MICRONUCLEI FORMATION, CELLULAR FRAGMENTATION, AND BLASTOMERE EXCLUSION

2017 ◽  
Vol 29 (1) ◽  
pp. 110 ◽  
Author(s):  
B. L. Daughtry ◽  
J. L. Rosenkrantz ◽  
N. Lazar ◽  
N. Redmayne ◽  
K. A. Nevonen ◽  
...  
Keyword(s):  
Rhesus Macaque ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Time Lapse ◽  
Confocal Imaging ◽  
Chromosomal Mosaicism ◽  
Human Embryos ◽  
Cleavage Stage ◽  
Chromosomal Breakage

A primary contributor to in vitro fertilization (IVF) failure is the presence of unbalanced chromosomes in pre-implantation embryos. Previous array-based and next-generation sequencing (NGS) studies determined that ~50 to 80% of human embryos are aneuploid at the cleavage stage. During early mitotic divisions, many human embryos also sequester mis-segregated chromosomes into micronuclei and concurrently undergo cellular fragmentation. We hypothesised that cellular fragmentation represents a response to mis-segregated chromosomes that are encapsulated into micronuclei. Here, we utilised the rhesus macaque pre-implantation embryo as a model to study human embryonic aneuploidy using a combination of EevaTM time-lapse imaging for evaluating cell divisions, single-cell/-fragment DNA-Sequencing (DNA-Seq), and confocal microscopy of nuclear structures. Results from our time-lapse image analysis demonstrated that there are considerable differences in the timing of the first and third mitotic divisions between rhesus blastocysts and those that arrested before this stage in development (P < 0.01; ANOVA). By examining the chromosome content of each blastomere from cleavage stage embryos via DNA-Seq, we determined that rhesus embryos have an aneuploidy frequency up to ~62% (N = 26) with several embryos exhibiting chromosomal mosaicism between blastomeres (N = 6). Certain blastomeres also exhibited reciprocal whole chromosomal gains or losses, indicating that these embryos had undergone mitotic non-disjunction early in development. In addition, findings of reciprocal sub-chromosomal deletions/duplications among blastomeres suggest that chromosomal breakage had occurred in some embryos as well. Embryo immunostaining for the nuclear envelope protein, LAMIN-B1, demonstrated that fragmented cleavage-stage rhesus embryos often contain micronuclei and that cellular fragments can enclose DNA. Our DNA-Seq analysis confirmed that cellular fragments might encapsulate whole and/or partial chromosomes lost from blastomeres. When embryos were immunostained with gamma-H2AX, a marker of chromatin fragility, we observed distinct foci solely in micronuclei and DNA-containing cellular fragments. This suggests that micronuclei may be ejected from blastomeres through the process of cellular fragmentation and, once sequestered, these mis-segregated chromosomes become highly unstable and undergo DNA degradation. Finally, we also observed that ~10% of embryos prevented cellular fragments or large blastomeres from incorporating into the inner cell mass or trophectoderm at the blastocyst stage (n = 5). Upon confocal imaging, multiple nuclei and intense gamma-H2AX foci were found in a large unincorporated blastomere in one of the blastocysts. Altogether, our findings demonstrate that the rhesus embryo responds to segregation errors by eliminating chromosome-containing micronuclei via cellular fragmentation and/or selecting against aneuploid blastomeres that fail to divide during pre-implantation development with significant implications for human IVF.

P–158 Assisted Hatching on D + 3 in order to facilitate trophectoderm biopsy in blastocyst for PGT-A is not advisable in all patients

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
P Belchin ◽  
Y Cabello ◽  
M Sanche. d. Burgos ◽  
J Guerrero ◽  
M D Riva ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Time Lapse ◽  
Fertilization Rate ◽  
Blastocyst Stage ◽  
Assisted Hatching ◽  
Culture Dish ◽  
Key Factor ◽  
Registration Number ◽  
Number Of Cycles

Abstract Study question Is it useful or beneficial to perform Assisted Hatching (AH) on D + 3 previously to biopsy for PGT-A on blastocyst stage on D + 5? Summary answer The routine use of AH on D + 3 to facilitate the embryo biopsy on D + 5 could negatively influence the development of the embryos to blastocyst stage. What is known already The blastocyst stage is the optimal stage for performing biopsies for PGT-A, which has been reported as a key factor determining the growing clinical application of this strategy worldwide. For trophectoderm (TE) biopsy, laser-assisted drilling is used to create a zona opening on D + 3 or D + 5 of development. The method of zona opening on D + 3 allows some of the TE cells to herniate during blastocyst formation and expansion, which facilitates the biopsy process. However, this method may result in herniation of inner cell mass cells instead of TE or maybe could affect the development of the embryo to blastocyst stage. Study design, size, duration A total of 100 PGT-A cycles were performed in 2019 and 2020. In 78 of them laser-assisted drilling was used to create a zona opening on D + 5 only in those embryos which arrived to blastocyst stage for TE biopsy (Group No-AH). In 22 cycles the same drilling was achieved on D + 3 in all embryos, independently of their quality (Group AH). The average of embryos per cycle in each group was 5 and 4.3 respectively. Participants/materials, setting, methods A total of 100 PGT-A cycles coming from 65 patients were studied. The average of the age of the patients was 40.83 (SD 3.45) in the group No-AH vs 42.18 (SD 3.42) in the Group AH (p = 0.108), so the age was not a determining factor for the development of the embryos. We analyzed by χ 2 test differences between groups on fertilization rates, number of embryos, development to blastocyst stage, euploidy and pregnancy rates. Main results and the role of chance The fertilization rate was 74.79% (No-AH group) and 68.53% (AH group) with no significative statistical differences (p = 0.12). In the No-AH group, the TE biopsy was performed on D + 5 in 63 cycles (81%). In the AH group, 41% of cycles didn’t reach the blastocyst stage, obtaining statistical differences between groups (p = 0.035). We found also significant differences in the number of cycles with biopsied blastocyst when we had 1 to 6 embryos/cycle on D + 3 between groups (p = 0.002), without obtaining any blastocyst to be diagnosed in 53% of the cycles in AH group vs 27% in No-AH group. When the number of embryos on D + 3 per cycle was &gt; 6, at least 1 embryo reached the blastocyst stage in both groups, although this number was higher in No-AH group. The rate of biopsied blastocysts was significantly higher in the No-AH group compared to the AH group (46.61 vs 34.69) with a p = 0.031. The rate of euploid embryos analyzed was 23.30% in the No-AH group compared to 29.41% in the AH group, although no significant differences were found (p = 0.44) between groups. In the No-AH group, a clinical pregnancy rate of 52.94% was obtained (n = 34) vs 50% in the AH group (n = 4) (p = 0.91). Limitations, reasons for caution We have recently started to perform AH on D + 3, so the number of cases is smaller than No-AH group. We use a time lapse incubator in all cases, so in the No-AH the culture dish is changed, disturbing the stable incubation environment, while in the other group it is not. Wider implications of the findings: The use of AH on D + 3 in order to facilitate the TE biopsy on D + 5 could affect negatively the development of the embryos to blastocyst stage. Its routine use should be avoided based on laboratory workload, mainly if the patient has less than 7 embryos at D + 3. Trial registration number Not applicable

Does Blastocyst Morphology Influence Live Birth Rate after Frozen-thawed Single Euploid Blastocyst Transfer?

2020 ◽  
Author(s):  
Na Li ◽  
Yichun Guan ◽  
Bingnan Ren ◽  
Yuchao Zhang ◽  
Yulin Du ◽  
...  
Keyword(s):  
Live Birth ◽  
Birth Rate ◽  
Inner Cell Mass ◽  
Statistical Significance ◽  
Cohort Analysis ◽  
Cell Mass ◽  
Live Birth Rate ◽  
Poor Quality ◽  
Significant Difference ◽  
Blastocyst Quality

Abstract Objective To determine whether the morphologic parameters of euploid blastocyst influence the live birth rate (LBR) following single frozen-thawed embryo transfer (FET) cycles? Methods A retrospective cohort analysis involving autologous single FET cycles after next generation sequencing (NGS) based preimplantation genetic testing for aneuploidy (PGT-A) by a large in vitro fertilization (IVF) center that was performed from June 2017 to September 2019.Women were divided into three age groups (< 30, 30–34 and ≥ 35 years old). The primary outcome measure was LBR. Outcomes were compared between different blastocyst quality (Good, Average and Poor), inner cell mass (ICM) grade (A and B), and trophectoderm (TE) grade (A, B and C). Results A total of 232 FET cycles were included, the live birth rate was 48.28%. In the youngest group (< 30 years old, n = 86), LBR were compared between cycles with various blastocyst quality (72.22% for good quality, 54.55% for average quality and 34.78% for poor quality; P = 0.019), ICM grade (70.59% for grade A and 42.03% for grade B; P = 0.035) and TE grade (85.71% for grade A,57.58% for grade B and 34.78 for grade C; P = 0.015). Nevertheless, either in the 30–34 years group (n = 99) or in the oldest group (≥ 35years, n = 47), LBR were also comparable between these subgroups, no significant difference was showed in blastocyst morphologic parameters and LBR (P > 0.05). Furthermore, in the similarly graded euploid blastocysts, there was also no statistical significance in LBR among different age subgroups (P > 0.05). Conclusions In women ≥ 30 years old, euploid blastocyst quality was not associated with the LBR in FET cycles, highlights the development competence of poor-quality euploid blastocysts.

Does Blastocyst Morphology Affect Live Birth Rate After Transfer of Single Euploid Embryo?

2021 ◽  
Author(s):  
Na Li ◽  
Yichun Guan ◽  
Bingnan Ren ◽  
Yuchao Zhang ◽  
Yulin Du ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Live Birth ◽  
Birth Rate ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Live Birth Rate ◽  
Poor Quality ◽  
Average Quality ◽  
Blastocyst Quality ◽  
Euploid Embryo

Abstract Background The aim of this study was to investigate whether the morphologic parameters of blastocyst influence the live birth rate (LBR) of euploid embryos transferred in subsequent single frozen-thawed embryo transfer (FET) cycles? Methods Women who received first preimplantation genetic testing for aneuploidy (PGT-A) and following underwent frozen-thawed single euploid blastocyst transfer cycles from June 2017 to May 2020 were divided into three age groups (< 30, 30–34 and ≥ 35 years). The primary outcome measure was LBR. Outcomes were compared between different blastocyst quality, inner cell mass (ICM) grade, trophectoderm (TE) grade and day of TE biopsy within the same age group. Results In the youngest group (< 30 years, n = 100), LBR were compared between cycles with various blastocyst quality (66.67% for good quality, 65.52% for average quality and 36.36% for poor quality; P = 0.013), ICM grade (61.11% for grade A and 51.22% for grade B; P = 0.466), TE grade (68.75% for grade A,65.00% for grade B and 36.30% for grade C; P = 0.012) and day of TE biopsy (65.38% for Day 5 and 39.58% for Day 6; P = 0.010). Similarly, in the 30–34 years group(n = 121) and the oldest group (≥ 35years, n = 58), LBR were also comparable between these subgroups, but no significant differences were seen between blastocyst morphologic grading and LBR (P > 0.05). Moreover, good quality (adjusted odds ratio [aOR] 3.30; 95% confidence interval [CI], 1.09 ~ 9.99; P = 0.035) and average quality (aOR 3.71; 95%CI, 1.25 ~ 11.01; P = 0.018) embryos were still yielded a significantly higher LBR than poor-quality embryos, TE grade B embryos were also associated with a statistically significantly higher LBR compared with TE grade C embryos (aOR 3.69;95%CI, 1.37 ~ 9.95; P = 0.010) after adjusting for the potential confounding factors. Conclusion Blastocyst quality and trophectoderm grading is a useful predictor of LBR in single frozen-thawed euploid embryo transfer cycles among women < 30 years old. However, these differences were not found in women older than 30 years.

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