P–131 Significance of the phenomenon of blastomere exclusion from compaction: Its relation to irregular cleavage, blastocyst development rate, and pregnancy rate

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Watanabe ◽  
M Tomida ◽  
S Suzuki ◽  
Y Matsuda ◽  
K Yoshikai ◽  
...  

Abstract Study question When does blastomere exclusion from compaction increase and what effect does it have on the embryo? Summary answer More blastomere were excluded from compaction in embryos with irregular cleavage, resulting in lower blastocyst development rates, but no decrease in pregnancy rates at transfer. What is known already It has been reported that many of the chromosome analysis results of blastomere excluded from compaction were aneuploid, and pointed out that this exclusion may be related to the repair of blastocyst euploidy, but the effect of the number of excluded blastomere has not been reported. Study design, size, duration This is a retrospective study of 578 embryos that developed into morula with time-lapse monitoring by EmbryoScope (Vitrolife) in 2018–2019. Participants/materials, setting, methods The target embryos were classified into two groups: embryos with normal first and second cleavage (normal cleavage group) and embryos with irregular cleavage (dynamics of one cell dividing into three or more cells), called “direct cleavage”, at either cleavage (DC group), and the number of blastomere excluded from compaction during morula formation was recorded and compared. The blastocyst development rate and single blastocyst transfer pregnancy rates of the two groups were compared. Main results and the role of chance There are 286 in the normal cleavage group and 292 in the DC group. The mean number of excluded blastomere was 0.76 and 3.55, respectively, which was significantly higher in the DC group (P < 0.01). Good blastocyst (Gardner classification 4 or higher) development rate was 84.5% (239/283) and 65.8% (181/275), respectively, and high grade blastocyst (Gardner classification BB or higher) development rate was 43.9% (105/239) and 14.9% (27/181) of them, both significantly higher in the normal cleavage group (P < 0.01). The single blastocyst transfer pregnancy rates were 31.6% (25/79) and 32.4% (11/34), and the miscarriage rates were 24.0% (6/25) and 27.3% (3/11), respectively, neither was there a significant difference between the two groups. So, direct cleavage increased the number of blastomere excluded from compaction, decreased the rate of morula to good blastocyst development and reduced blastocyst grade, but did not affect blastocyst transfer pregnancy rate and miscarriage rate. Limitations, reasons for caution Please note that all target embryos must have developed into morula or larger (embryos that did not develop into morula will not be included in the study). Wider implications of the findings: Severe chromosomal aberrant blastomeres formed by direct cleavage were excluded from compaction, and the blastocyst development rate decreased due to a decrease in the amount of viable cells, but it is suggested that this blastomere exclusion mechanism is not related to euploidy after blastocyst development. Trial registration number Not applicable

2020 ◽  
Vol 35 (11) ◽  
pp. 2478-2487
Author(s):  
Jiayi Wu ◽  
Jie Zhang ◽  
Yanping Kuang ◽  
Qiuju Chen ◽  
Yun Wang

Abstract STUDY QUESTION Does cell number on Day 3 have an impact on pregnancy outcomes in vitrified-thawed single blastocyst transfer cycles? SUMMARY ANSWER A low Day 3 cell number (≤5 cells) was independently associated with decreased live birth rate (LBR) during single blastocyst transfer cycles in young women. WHAT IS KNOWN ALREADY Day 3 cell number is an effective predictor of IVF success rates when transferring cleavage stage embryos. However, the association between Day 3 blastomere number and pregnancy outcomes after blastocyst transfer is still unknown. STUDY DESIGN, SIZE, DURATION A retrospective cohort study of 3543 patients who underwent frozen-thawed single blastocyst transfers from January 2013 to June 2018 at a tertiary-care academic medical center. PARTICIPANTS/MATERIALS, SETTING, METHODS Patients were grouped into six groups according to the Day 3 cell number: ≤4 cells, 5 cells, 6 cells, 7 cells, 8 cells and >8 cells. The primary outcome measure was LBR. A logistic regression analysis was performed to explore the independent association between Day 3 blastomere number and LBR after adjustment for some potential confounders. MAIN RESULTS AND THE ROLE OF CHANCE In women <35 years old, the LBR varied significantly according to Day 3 cell number, with the rate of 31.2%, 34.4%, 41.9%, 45.1%, 48.1% and 48.2% for the ≤4-cell, 5-cell, 6-cell, 7-cell, 8-cell and >8-cell groups, respectively (P < 0.001). This significant difference was also observed in the high- and low-quality blastocyst subgroups of young women. However, for women ≥35 years old, the rate of live birth was similar between groups. Furthermore, after accounting for confounding factors, the LBR was significantly decreased in the ≤4-cell (adjusted odds ratio (aOR): 0.62, 95% CI: 0.48–0.80, P < 0.001) and 5-cell (aOR: 0.73, 95% CI: 0.57–0.92, P = 0.009) groups as compared to the 8-cell group. Likewise, the blastocysts arising from ≤4-cell (aOR: 0.73, 95% CI: 0.57–0.93, P = 0.010) or 5-cell (aOR: 0.77, 95% CI: 0.61–0.97, P = 0.024) embryos were associated with lower clinical pregnancy rate than those from 8-cell embryos. No significant differences were observed in biochemical pregnancy rate and miscarriage rate. LIMITATIONS, REASONS FOR CAUTION A limitation of the current study was its retrospective design. Future prospective studies are needed to confirm our findings. WIDER IMPLICATIONS OF THE FINDINGS Our observations suggested that a low Day 3 cell number was related to decreased LBR after blastocyst transfer in young women, which provided vital information for clinicians in selecting blastocyst during IVF treatment. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the National Natural Science Foundation of China (NSFC) (31770989 to Y.W.; 81671520 to Q.C.) and the Shanghai Ninth People’s Hospital Foundation of China (JYLJ030 to Y.W.). The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER N/A.


2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Weijue Su ◽  
Jialing Xu ◽  
Samuel Kofi Arhin ◽  
Chang Liu ◽  
Junzhao Zhao ◽  
...  

Objective. To investigate the feasibility and clinical outcome of the all-blastocyst-culture and single blastocyst transfer strategy in women aged ≥35 years. Methods. A retrospective analysis of patients aged ≥35 years undergoing IVF/ICSI was performed from January 2017 to April 2019 in the reproductive center of the Second Affiliated Hospital of Wenzhou Medical University. A total of 155 cases treated with ovarian hyperstimulation by prolonged protocol and implemented single (84 cases) or double (71 cases) blastocyst transfer were collected. Then, patients were further divided into <38 yr. group and ≥38 yr. group, and the laboratory and clinical outcomes were compared between the groups. Results. The double-blastocyst-transfer (DBT) group showed higher clinical pregnancy rate and multiple pregnancy rate and lower neonatal birth weight than those in the single-blastocyst-transfer (SBT) group (P<0.05). However, there were no statistically significant differences between the groups in the embryo implantation rate, biochemical pregnancy rate, miscarriage rate, preterm delivery rate, and term birth rate. For patients<38 yr., SBT significantly reduced the multiple pregnancy rate and increased the neonate birth weight without significant reduction in the clinical pregnancy rate. While in the ≥38 yr. group, there are no differences in pregnancy outcomes between SBT and DBT. Logistic regression analysis showed that the number of MII oocytes was positively correlated with the live birth rate (OR=1.18) and negatively correlated with the miscarriage rate (OR=0.844), suggesting that elderly patients with relatively normal ovarian reserve would obtain better prospect in pregnancy. The number of fetal heart beat in pregnancy was negatively correlated with the live birth rate (OR=0.322) and positively correlated with the preterm birth rate (OR=7.16). Conclusion. The strategy of all-blastocyst-culture and single blastocyst transfer is feasible, safe, and effective for elderly patients with normal ovarian reserve, which would reduce the multiple pregnancy rate.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Garcia-Faura ◽  
B Marques ◽  
V Montalvo ◽  
F Garcia ◽  
M Lopez-Teijon

Abstract Study question Looking at time-lapse images of own embryos during the five days of culture, as a patient’s active behaviour, may help to increase pregnancy rates? Summary answer The number of visual connections to live blastocyst development images of own embryos positively correlates to patient’s pregnancy rates after fresh single blastocyst transfer. What is known already In human reproduction, despite the evidence that infertility itself and reproductive treatments can produce anxiety and stress, the impact of emotions on pregnancy outcomes is unknown and probably underestimated. The interaction between psyche, nervous, immune and endocrine systems (PNIE) is well known and has been applied to understand and treat different pathological conditions. Patient’s active behaviour during IVF treatments appears to be a positive strategy to decrease women’s anxiety and stress, thus to increase fertility quality of life and pregnancy outcomes. Study design, size, duration Retrospective comparative study of 934 patients undergoing fresh IVF cycles during three years. To avoid any bias related to oocyte and embryo factors, we only included egg donation cycles in the study, and ≥3BB (Gardner score) single embryo transfers. All embryo cultures were performed in time-lapse incubator and patients could connect on-line to their embryo’s images before single blastocyst transfer. We evaluated the impact of visual connections to live embryo images on pregnancy rates. Participants/materials, setting, methods On day one of embryo culture, consenting patients received an individualized and secure link allowing them to connect on-line to images of their own embryos any time during the five days of culture. Patients were divided in five groups depending on the number of on-line visualisations (A = 0; B = 1–10; C = 11–20; D = 21–30; E &gt; 30). Pregnancy and clinical pregnancy rates were compared between the five groups. Chi-square test for a large contingency table was performed to compare all groups Main results and the role of chance The distribution of patients in the five groups, based on number of visualizations, resulted as follows: 287 in group A; 328 in group B; 156 in group C; 80 in group D; 83 in group E. The five groups were homogeneous and there were no statistically significant differences in recipient’s ages (A 42.8±3.9 years; B 41.9±3.9 years; C 41.8±4.5 years; D 42.8±4.1 years; E 41.9±4.4 years) and in donor’s ages (A 25.5±4.3 years; B 26.9±4.4 years; C 27.1±4.2 years; D 26.4±4.5 years; E 26.5±4.1 years) among the five groups. We observed a progressive positive trend between the number of on-line visualisations and pregnancy rates, reaching statistical significance for group E (&gt;30 visualizations) compared to the others groups. In group E, pregnancy rates and clinical pregnancy rates per fresh single blastocyst transfer were respectively 72.3% and 65.1%, significantly higher when compared to the others groups (p &lt; 0.01, p &lt; 0.001): group A 61% and 50.9%, group B 63.1% and 56.1%, group C 64.1% and 55.1%, group D 65% and 53.8%. Limitations, reasons for caution Time-lapse incubator is needed as well as a strong informatics setting to ensure secure and personalised on-line connection to each patient at any time during the five days of culture. Endocrine and endometrial biomarkers should be proposed and evaluated in further clinical trials to better understand these results. Wider implications of the findings: Repeated on-line visualization of own embryos images before fresh blastocyst transfer enhances pregnancy rates in IVF cycles. We propose that repeated visual stimuli of images produces a positive emotional connection between patients and their own developing embryos, thus reducing anxiety and enhancing recipient’s endometrial receptivity. Trial registration number Not applicable


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yuxia He ◽  
Shiping Chen ◽  
Jianqiao Liu ◽  
Xiangjin Kang ◽  
Haiying Liu

Abstract Background High-quality single blastocyst transfer (SBT) is increasingly recommended to patients because of its acceptable pregnancy outcomes and significantly reduced multiple pregnancy rate compared to double blastocyst transfer (DBT). However, there is no consensus on whether this transfer strategy is also suitable for poor-quality blastocysts. Moreover, the effect of the development speed of poor-quality blastocysts on pregnancy outcomes has been controversial. Therefore, this study aimed to explore the effects of blastocyst development speed and morphology on pregnancy and neonatal outcomes during the frozen embryo transfer (FET) cycle of poor-quality blastocysts and to ultimately provide references for clinical transfer strategies. Methods A total of 2,038 FET cycles of poor-quality blastocysts from patients 40 years old or less were included from January 2014 to December 2019 and divided based on the blastocyst development speed and number of embryos transferred: the D5-SBT (n = 476), D5-DBT (n = 365), D6-SBT (n = 730), and D6-DBT (n = 467) groups. The SBT group was further divided based on embryo morphology: D5-AC/BC (n = 407), D5-CA/CB (n = 69), D6-AC/BC (n = 580), and D6-CA /CB (n = 150). Results When blastocysts reach the same development speed, the live birth and multiple pregnancy rates of DBT were significantly higher than those of SBT. Moreover, there was no statistical difference in the rates of early miscarriage and live birth between the AC/BC and CA/CB groups. When patients in the SBT group were stratified by blastocyst development speed, the rates of clinical pregnancy (42.44 % vs. 20.82 %) and live birth (32.35 % vs. 14.25 %) of D5-SBT group were significantly higher than those of D6-SBT group. Furthermore, for blastocysts in the same morphology group (AC/BC or CA/CA group), the rates of clinical pregnancy and live birth in the D5 group were also significantly higher than those of D6 group. Conclusions For poor-quality D5 blastocysts, SBT can be recommended to patients because of acceptable pregnancy outcomes and significantly reduced multiple pregnancy rate compared with DBT. For poor-quality D6, the DBT strategy is recommended to patients to improve pregnancy outcomes. When blastocysts reach the same development speed, the transfer strategy of selecting blastocyst with inner cell mass “C” or blastocyst with trophectoderm “C” does not affect the pregnancy and neonatal outcomes.


2011 ◽  
Vol 23 (1) ◽  
pp. 211
Author(s):  
Y. S. Park ◽  
H. D. Park

In conventional in vitro embryo production, oocytes are obtained from various ovaries, and donor characteristics, such as pedigree type, meat quality, or meat quantity, are not considered. The aim of this study was to examine the effects of pedigree type and meat quality and quantity of slaughterhouse donor cows on oocyte recovery, embryo development, and pregnancy rates. The ovaries of individual Korean native cows were obtained from a slaughterhouse. The ear tag numbers were obtained 24 h after slaughter, and the donor data were compared according to the pedigree type (no registration to third bloodline registration), meat quantity according to the percentage of carcass weight (more than 69%, 66 to 69%, and below 66%; Grades A, B, and C, respectively), and meat quality according to the marbling grade (marbling grade 8 to 9, 6 to 7, 4 to 5, 2 to 3, and 1; Grades 1++, 1+, 1, 2, and 3, respectively). There were 390 donors in total, and each group had 30 donors. Recipients (Holstein heifers; n = 222) were synchronized with a progesterone device, and a single blastocyst was transferred nonsurgically on Day 7 from the onset of oestrus. Data on embryo development were compared by Duncan’s multiple range tests, and pregnancy rate was analysed by chi-square test. Values of P < 0.05 were considered to indicate a significant difference. No significant difference was detected in the average number of oocytes recovered (17.5 ± 2.9 to 22.9 ± 1.2%) or blastocyst development (16.4 ± 2.6 to 19.5 ± 3.4%) from cows of different pedigrees. A higher number of oocytes were recovered from Grade A (22.9 ± 1.0) than from Grade B (19.8 ± 0.7; P < 0.05) cows. However, the meat quantity grade had no significant effect on blastocyst development (14.3 ± 1.2 to 15.0 ± 0.8%). A greater number of oocytes were recovered from the Grade 1++ (25.0 ± 1.6), 1+ (27.7 ± 2.4), and 2 (23.6 ± 1.5) groups than from the Grade 3 group (19.7 ± 1.0; P < 0.05). The developmental rate to the blastocyst stage was significantly higher in the Grades 2 and 3 groups (22.1 ± 2.2 and 19.4 ± 1.7) than in the Grade 1++, 1+, and 1 groups (9.0 ± 1.2 to 13.8 ± 2.0). Pregnancy rate was not affected by pedigree type, meat quantity, or meat quality (41.7 to 58.6%). We concluded that 1) pedigree type had no effect on oocyte recovery and embryo development; 2) meat quantity affected oocyte recovery but did not affect embryo development; 3) high meat quality (Grades 1++ and 1+) resulted in greater oocyte recovery but lower embryo development; and 4) pregnancy rate remained unaffected by pedigree type, meat quantity, and meat quality. This research was supported by the Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.


2015 ◽  
Vol 27 (1) ◽  
pp. 194
Author(s):  
S. Saugandhika ◽  
H. N. Malik ◽  
S. Saini ◽  
V. Sharma ◽  
S. Bag ◽  
...  

Interferon tau (IFN-tau) is known as maternal pregnancy recognition factor in ruminants. IFN-tau not only acts as a signalling molecule of pregnancy recognition but also performs various functions for successful implantation and pregnancy establishment. The aim of the present study was to produce recombinant buffalo interferon-tau (BuIFN-Tau) and observe if it has any effect on in vitro embryo development. The BuIFN-Tau gene was obtained through polymerase chain reaction (PCR) from hatched buffalo blastocysts and was cloned into pJET cloning vector. Screening of the recombinant colonies gave 8 distinct buffalo IFN-tau isoforms, out of which the predominant buffalo IFN-t tau1 isoform (gene bank accession number JX481984), was subcloned into expression vector pET22b without signal sequence. The recombinant plasmid was induced to express the recombinant protein by isopropyl b-D-1-thiogalactopyranoside. Analysis of the products of recombinant BuIFN-tau without signal sequence by SDS–PAGE revealed a new 20-kDa protein coinciding with the molecular weight of IFN-tau as reported earlier in literature. The purified recombinant BuIFN-tau was confirmed by Western blot using anti-HIS antibody and was subjected to three steps of large-scale purification using HIS affinity chromatography, anion exchange chromatography, and gel filtration chromatography. Finally, a relatively pure histidine-tagged recombinant protein, which had a purity of at least 90%, was generated as confirmed through SDS. The concentration of recombinant BuIFN-tau was 1 mg mL–1 by Bradford assay. The purified recombinant BuIFN-tau was used as supplement of the culture medium for IVF early buffalo embryos at the following concentrations: control, 1, 2, and 4 µg mL–1. Sixty oocytes each in 4 groups (with 20 oocytes/drop in three replicates for each group) were used for in vitro maturation. After 24 h, the matured oocytes were incubated with in vitro capacitated sperm cells for 18 h; thereafter, the presumptive zygotes were cultured in IVC medium supplemented with 0, 1, 2, or 4 µg mL–1 of the purified recombinant BuIFN-tau. The experiment was repeated 3 times. The data were analysed using SYSTAT 7.0 (SPSS Inc., Chicago, IL, USA) after arcsin transformation of percentage values. The differences were analysed by one-way ANOVA followed by Fisher's least significant difference test. Out of 3 concentrations of recombinant BuIFN-tau, the 2 µg mL–1 concentration significantly promoted the rate of blastocyst development, 45.55% against 31.1% (control; P < 0.01). Blastocyst development rate for low and high concentrations was 29.97% and 10.18% respectively. It is concluded that the addition of 2 µg mL–1 of recombinant BuIFN-tau enhances the blastocyst development rate in buffalo, and hence there is some evidence that BuIFN-tau has not only a role in maternal recognition of pregnancy but also in embryonic development.


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