Does the sex ratio of singleton births after frozen single blastocyst transfer differ according to blastocyst morphology?

Abstract Purpose: To investigate the associations between morphological blastocyst parameters and the sex ratio (male:female) of singleton live births resulting from single-blastocyst frozen embryo transfer (FET) cycles. Methods: This retrospective analysis included 1210 couples who underwent single-blastocyst FET of warmed day 5 or 6 blastocysts and achieved a singleton live birth between January 2015 and February 2019. The sex ratios in the different blastocyst groups were compared using the chi-square test. The associations of morphological blastocyst parameters, including the blastocoel expansion, inner cell mass and trophectoderm grades, with the sex ratios of live births were analyzed using multiple regression models. Results: The included female patients had an average age and body mass index of 30.5±4.5 years and 23.6±3.2 kg/m2, respectively. blastocyst transfers occurred on day 5 in 783 cases (64.7%) and day 6 in 427 cases (35.3%). Among the day 5 FET cycles, 55.4% of resulting infants were male and 44.6% were female, while among the day 6 cycles, 54.6% of infants were male and 45.4% were female ( P =0.074). Blastocysts quality was assessed according to morphological parameters, which was divided into high-quality (AA,AB,BB,BC) and poor-quality(AC,BC). High-quality blastocyst transfer was associated with a significantly higher sex ratio (60%, 383/638) relative to poor-quality transfer (49.7%, 284/572) ( P< 0.001). The sex ratio of births resulting from blastocyst transfer differed significantly with respect to the trophectoderm grade ( P <0.001). After adjusting for confounders, a grade B trophectoderm was significantly associated with a higher sex ratio among singleton live births (odds ratio: 0.591, 95% confidence interval: 0.463–0.756, P <0.001; reference: grade C trophectoderm). In contrast, the characteristics of blastocoel expansion and the score of the inner cell mass were not significantly associated with the sex ratio among singleton live births. Conclusions: The sex ratio among singleton live births is affected by the quality of blastocysts. A grade B trophectoderm is associated with a higher probability that a single-blastocyst FET cycle will result in a male infant.

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

10.21203/rs.3.rs-1011865/v1 ◽

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.

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

10.21203/rs.3.rs-149923/v1 ◽

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.

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Cytoplasmic String Between ICM and mTE Is a Positive Predictor of Clinical Pregnancy and Live Birth Outcomes in Elective Frozen-thawed Single Blastocyst Transfer Cycles: a Time-lapse Study

10.21203/rs.3.rs-122470/v1 ◽

2020 ◽

Author(s):

Bing-Xin Ma ◽

Lei Jin ◽

Bo Huang

Keyword(s):

Birth Outcomes ◽

Live Birth ◽

Inner Cell Mass ◽

Cell Mass ◽

Time Lapse ◽

Clinical Pregnancy ◽

Blastocyst Transfer ◽

Single Blastocyst Transfer ◽

First Time ◽

Present Group

Abstract Background: In this study, we aim to investigate whether cytoplasmic string between inner cell mass (ICM) and mural trophectoderm (mTE) is a positive predictor of clinical pregnancy and live birth outcomes.Methods: 1,267 elective frozen-thawed single blastocyst transfer (eSBT) cycles cultured in time-lapse incubation system from January 2018 to May 2019 were involved in the study. Blastocysts were grouped according to the appearance of cytoplasmic strings between ICM and mTE cells, and identified as “Present” and “Absent” groups. In Present group, they were further categorized according to the quantity of cytoplasmic strings between ICM and mTE cells. Clinical pregnancy and live birth outcomes of blastocysts were used to evaluate the effect of cytoplasmic strings between ICM and mTE.Results: The baseline demographic and laboratory features were similar between the Present and Absent groups of cytoplasmic strings between ICM and mTE (P>0.05). According to the time-lapse analysis, cytoplasmic strings between ICM and mTE were more visible among good quality blastocysts. Furthermore, blastocysts with cytoplasmic strings showed a higher clinical pregnancy and live birth rates (P<0.05), and no significant differences were observed in abortion rate and birth weight (P>0.05).Conclusions: Although the previous conclusions of cytoplasmic strings were controversial, the present time-lapse analysis provides the evidence for the first time that cytoplasmic strings between ICM and mTE cells would be a positive predictor of clinical pregnancy and live birth outcomes in elective frozen-thawed single blastocyst transfer cycles.

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

10.21203/rs.3.rs-131940/v1 ◽

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.

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

10.21203/rs.3.rs-787954/v1 ◽

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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The Influence of Day 2 Blastomere Symmetry on Blastocyst Grade and Ploidy Status

Fertility & Reproduction ◽

10.1142/s2661318219500117 ◽

2019 ◽

Vol 01 (02) ◽

pp. 115-118

Author(s):

Diana Chieh Xing Tain ◽

Michelle Sheng Rong Lim ◽

Bee Lian Ng ◽

Elizabeth Hammond ◽

Pak Seng Wong

Keyword(s):

Inner Cell Mass ◽

Formation Rate ◽

Cell Mass ◽

Time Lapse ◽

Blastocyst Stage ◽

Chi Square ◽

Cell Stage ◽

Cell Asymmetry ◽

Time Lapse Imaging ◽

Ploidy Status

Previous studies have suggested that aneuploidy rates are co-related with cell asymmetry at the cleavage stage. A retrospective study was carried out to determine the significance of blastomere symmetry at the 4-cell stage on blastocyst grade and ploidy status. 732 Day 5/6 blastocysts from 191 patients undergoing Pre-implantation Genetic Testing for Aneuploidy were analysed with time-lapse imaging (Embryoscope, Vitrolife) during 2017. Blastomere symmetry was measured at the first image of 4-cells on Day 2 by tabulating the mean diameter of 2 lines drawn perpendicularly on each blastomere. Symmetry was defined as the blastomere diameter difference of [Formula: see text] 25%. Trophectoderm (TE) biopsy was performed on Day 5/6 followed by chromosomal evaluation using Next Generation Sequencing (VeriSeq Protocol, Illumina). Blastocyst grade was classified as either “Good” (inner cell mass (ICM) and TE, AA respectively), “Fair/Good” (AB, BA), “Fair” (BB) and “Poor” (early blastocyst grade 2 or TE grading of C). The significance of blastomere symmetry on blastocyst grade and ploidy status was measured using chi-square tests. There was no significance difference in resulting blastocyst quality for symmetrical and asymmetrical embryos (Table 1: p [Formula: see text] 0.10). Furthermore, there was no significance difference in the euploid rate (42.5% vs. 45.3%) or mosaic rate (22.1% vs. 16.2%) between symmetrical and asymmetrical embryos (p [Formula: see text] 0.24). In conclusion, the presence of asymmetrical blastomeres at the 4-cell stage do not impact the good quality blastocyst formation rate and euploidy rate for embryos that progress into blastocysts. However, this study excludes embryos that do not develop to the blastocyst stage and those with erratic division patterns, direct cleavage and reverse cleavage on Day 2, both of which have potential to influence ploidy result. Asymmetrical 4-cell embryos have the potential for high quality euploid blastocyst progression and can be considered for day 2 embryo transfer in the absence of symmetrical 4-cell embryos.

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Characterizing meiotic and mitotic errors in the inner cell mass and trophectoderm of poor quality preimplantation embryos

Fertility and Sterility ◽

10.1016/j.fertnstert.2019.07.1383 ◽

2019 ◽

Vol 112 (3) ◽

pp. e248

Author(s):

Christine Briton-Jones ◽

Lucky Sekhon ◽

Ethan Ellis ◽

Joseph A. Lee ◽

Eric E. Schadt ◽

...

Keyword(s):

Inner Cell Mass ◽

Cell Mass ◽

Poor Quality ◽

Preimplantation Embryos

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

10.21203/rs.3.rs-534083/v1 ◽

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 ◽

Age Groups ◽

Live Birth Rate ◽

Poor Quality ◽

Preimplantation Genetic Testing ◽

Blastocyst Quality

Abstract Background: The aim of this study was to investigate whether the morphologic parameters of euploid blastocyst influence the live birth rate (LBR) following single frozen-thawed embryo transfer (FET) cycles?Methods: Women who undergone first preimplantation genetic testing for aneuploidy (PGT-A) and following received single FET cycles from June 2017 to September 2019 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 among 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: 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 between blastocyst morphologic parameters and LBR (P>0.05). Conclusion: 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.

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Cytoplasmic String Between ICM and mTE Is a Positive Predictor of Clinical Pregnancy and Live Birth Outcomes in Elective Frozen-thawed Single Blastocyst Transfer Cycles: A Time-lapse Study

10.21203/rs.3.rs-144098/v1 ◽

2021 ◽

Author(s):

Bingxin Ma ◽

Lei Jin ◽

Bo Huang

Keyword(s):

Birth Outcomes ◽

Live Birth ◽

Inner Cell Mass ◽

Cell Mass ◽

Time Lapse ◽

Clinical Pregnancy ◽

Blastocyst Transfer ◽

Single Blastocyst Transfer ◽

First Time ◽

Present Group

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148 MICE BLASTOCYST RECONSTRUCTION BY AGGREGATION OF INNER CELL MASS WITH TROPHECTODERM

Reproduction Fertility and Development ◽

10.1071/rdv25n1ab148 ◽

2013 ◽

Vol 25 (1) ◽

pp. 222

Author(s):

I. P. Emanuelli ◽

C. P. Godoi ◽

M. P. M. Mancini ◽

C. M. Barros ◽

M. F. G. Nogueira

Keyword(s):

Present Model ◽

Inner Cell Mass ◽

Cell Mass ◽

Cell Aggregation ◽

Control Group ◽

Control Groups ◽

Chi Square ◽

Trophectoderm Cells ◽

And Control

In order to reduce fetal losses of bovine clones the replacement of trophectoderm (TE) by microinjection of immunosurgically isolated inner cell mass (ICM) was proposed. It was reported that those techniques could damage both ICM and TE. An alternative to decrease this problem would be the technique of cell aggregation by approximation. Currently it has been used only in pre-compaction embryos. The aims of this study were to evaluate the effectiveness to aggregate post-compaction embryos and to verify the feasibility of reconstructed blastocysts production by ICM and TE approximation. Mice (Swiss Webster, SW; and C57BL/6/EGFP, EGFP) were superovulated to obtain embryos 2.5 to 3.5 days post-coitus. In both control groups (CG; exp. 1 and 2), whole embryos from 8 cells to early morula stages and without zona pellucida were aggregated. In all experiments, bisection was performed with a blade controlled by micromanipulator to produce two fragments according to the requirements of each experiment (i.e. demi embryos in Exp. 1 and Exp. 2 or TE fragment and whole ICM in Exp. 3). The joined fragments were in vitro cultured in microwell (300 µm diameter and based on well of the well system) filled with 400 µL of KSOMaa, covered with mineral oil by 24 h (incubation 37°C, 5% of CO2 and saturated humidity). Embryonic aggregation rate (AR) was evaluated by detection of a single and cohesive cell mass (CG from Exp. 1 and Exp. 2) or by the presence of a typical morphologically blastocyst (all the remaining groups). Only to validate AR evaluation, 25% of the aggregation attempts were performed with distinct phenotype embryos (EGFP and SW) and they could be tracked under epifluorescence. In Exp. 1, the aggregation by approximation was tested to evaluate embryonic stages: 2 demi-blastocysts (2DB; n = 28), demi-morula and demi-blastocyst (DMDB; n = 20) and control group (CG; n = 25). In Exp. 2, an adhesive agent (phytohemagglutinin; PHA) and fragment increasing were used in the groups: 2DB (n = 24), 4 demi-blastocysts (4DB, that is four half-blastocyst) and CG (n = 22). After aggregation validation (Exp. 1 and Exp. 2), blastocyst reconstruction by the approximation of ICM and TE fragments (with PHA presence) was tested on groups: 1 TE fragment and 1 ICM (TI; n = 48) and 2 TE fragments and 1 ICM (2T1I; n = 17). The rates were analyzed by chi-square and Fisher’s exact tests (significance of 5%). In experiments 1 (3.6; 15.0; and 60.0%, respectively to 2DB, DMDB, and CG) and 2 (8.3; 36.4; and 77.3%, respectively to 2DB, 4DB, and CG), the AR differed among groups with exception of 2DB and DMDB (exp. 1; P > 0.05). In Exp. 3, there was no difference on the AR between TI and 2T1I (27.1 and 29.4%, respectively). Despite the low adhesion potential of the trophectoderm cells, is feasible to produce chimeric embryos by aggregation. We infer that the use of an adhesive agent could increase AR maintaining the contact of the embryonic fragments. The proposed technique was practical and effective for blastocyst reconstruction. Additionally, the present model established in mice is about to be tested in cattle for further validation. Financial support: FAPESP.

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