Embryo quality and chromosomal abnormality in embryos from couples undergoing assisted reproductive technology using preimplantation genetic screening

Objective:To detect common chromosomal aneuploidy variations in embryos from couples undergoing assisted reproductive technology and preimplantation genetic screening and their possible associations with embryo quality.Methods:In this study,359 embryos from 62 couples were screened for chromosomes 13,21,18,X,and Y by fluorescence insitu hybridization.For biopsy of blastomere,a laser was used to remove a significantly smaller portion of the zona pellucida.One blastomere was gently biopsied by an aspiration pipette through the hole.After biopsy,the embryo was immediately returned to the embryo scope until transfer.Embryo integrity and blastocyst formation were assessed on day 5.Results:Totally,282 embryos from 62 couples were evaluated.The chromosomes were normal in 199(70.57%)embryos and abnormal in 83(29.43%)embryos.There was no significant association between the quality of embryos and numerical chromosomal abnormality(P=0.67).Conclusions:Embryo quality is not significantly correlated with its genetic status.Hence,the quality of embryos determined by morphological parameters is not an appropriate method for choosing embryos without these abnormalities.

Relationship between Pronuclear Scoring and Embryo Quality and Implantation Potential in IVF-ET

To assess the relationship between pronuclear scoring and day-3 embryo quality and pregnancy outcome and to determine the Clinical value of pronuclear stage scoring system in human in vitro fertilization-embryo transfer （IVF-ET） program, a pronuclear scoring system was used to score zygotes 16-20 h after insemination during conventional IVF or intracytoplasmic sperm injection （ICSI）. The embryos were classified into groups Z1, Z2, Z3 and Z4, Comparisons were made of the rates of arrested embryos and excellent embryos on day 3. Comparisons of pregnancy outcome were made only in those patients in whom cohorts of similarly Z-scored embryos were transferred, The results showed that there were less arrested embryos and more excellent embryos on day 3 in groups Z1 and Z2 than those in group Z3 and Z4, More embryos arrested and less excellent embryos developed in group Z4 than group Z3. The clinical pregnancy rates resulting from the transfer of single pronuclear score homologous embryo types were similar among groups Z1, Z2 and Z3. Implantation rates of group Z1 were higher （P〈0.05） than that of group Z3, These findings suggests that pronuclear scoring can predict developmental ability on day 3 and implantation potential. A evaluation that combines the Z-score and day 3 embryo morphology is useful in the determination of the most viable embryos and the number of embryos for transfer.

Free radical scavenging window of infertile patients with polycystic ovary syndrome： correlation with embryo quality

The activity of free radicals in follicular fluid was related to ovarian responsiveness, in vitro fertilization （IVF）, and embryo transfer success rate. However, studies analyzing the relationship between the free radical scavenging capacity and embryo quality of infertile women with polycystie ovarian syndrome （PCOS） were lacking. The aim of this study was to evaluate the relationship between the free radical scavenging window of women with PCOS and their embryo quality. The free radical scavenging capacity of follicular fluid from women with PCOS was determined by a,a-diphenyl-b-picrylhydrazyl （DPPH）, 2,2-azinobis （3-ethylbenzthiazoline-6- sulphonic acid） assay, superoxide radical, and reactive oxygen species （ROS） assay. In the DPPH and ROS assays, the follicular fluid from grades I and II embryos was significantly higher than the follicular fluid from grades III and IV embryos. The lower control limit of DPPH radical scavenging capacity and upper control limit of ROS level were 13.2% and 109.0 cps, respectively. The calculated lower control limit and upper control limit were further confirmed in the follicular fluid of embryos of all grades. These cut-off values of free radical scavenging activity of follicular fluid could assist embryologists in choosing the development of embryos in PCOS patients undergoing IVF.

Effect of Semen Quality on the Embryo Development

To investigate the influences of sperm quality on the zygotes and embryos development, as the role of the paternal factor in early human embryogenesis is gaining more attention because of the application of techniques such as intracytoplasmic sperm injection （ICSI） ior the treatment of men infertility. 136 infertility couples with men factors （Group Ⅰ） were included from May 2002 to January 2004. One hundred and seventy-two infertility couples with tube factors （Group Ⅱ） served as controls. The sperm parameters, geminates and embryos quality, implantation rate and pregnant rate in both groups were analyzed. It was found that there was no significant differences in the number of oocytes retrieved, the fertilization rate and number of embryos transferred between two groups. Sperm concentration, percentage of motile sperm and percentage of sperm with normal morphology were significantly lower in group Ⅰ than in group Ⅱ （P〈0.01）. The proportion of good quality zygotes and good quality embryos were significantly lower in,（he male infertility group than in the tubal disease group （P（0.05）. Implantation rate and pregnancy rate were similar in two groups. It was concluded that spermatozoa is involved in the embryo quality, even in the early stages of development, which limited the treatment potency of IVF procedure.

Granulosa Cell Biomarkers to Predict Oocyte and Embryo Quality in Assisted Reproductive Technology

With the development of human assisted reproductive technology(ART),an objective,accurate,and non-invasive method to assess the quality and viability of oocytes and embryos remains one of the most significant goals.Granulosa cells(GCs)play an essential role in oocyte development.GCs can differentiate into mural GCs(MGCs)and cumulus cells(CCs)under the influence of oocytes.MGCs promote the growth and development of follicles by secreting cytokines and steroid hormones.Simultaneously,CCs can form cumulus-oocyte complexes to communicate with oocytes through gap junctions and promote oocyte growth and maturation.Seeking suitable biomarkers in GCs provides a direction for the non-invasive assessment of oocyte and embryo abilities during ART procedures.To date,only a few studies have investigated potentially effective GC biomarkers during ART processes,such as the apoptosis of GCs,transcriptomic characteristics of GCs,quality and quantity of mitochondria in GCs,and telomere length of such cells.These are potential reference indices for screening high-quality oocytes and embryos.Independent studies on MGCs and CCs can provide more effective results.Although there is scope for optimization and improvement,the results have become increasingly accurate with the constant advances in technology.Due to the heterogeneity of the study population and technical limitations,clinical tests for GCs cannot be performed as part of routine tests,but their prospects are promising.This article reviews the biomarkers that have been studied in MGCs and CCs.

Pregnancy Outcomes for Day 5 Versus Day 6 Single Frozen-thawed Blastocyst Transfer with Different Qualities of Embryos: A Large Matched-cohort Study

Objective This study aimed to determine whether the day of blastocyst expansion affects pregnancy outcomes in frozen-thawed blastocyst transfer(FBT)cycles.Methods A retrospective match-cohort study was conducted.Patients who underwent blastocyst transfer in frozen-thawed cycles at day 5 or 6 were matched for potential confounding factors.A total of 2207 matched pairs of FBT cycles were included from January 2016 to December 2019 in our Reproductive Medicine Center.Results The clinical pregnancy rate(CPR)and live birth rate(LBR)were significantly increased in day 5 blastocyst transfers when compared to day 6 blastocyst transfers,in terms of the same embryo quality.For FBT cycles with good-quality embryo,the CPR at day 5 and 6 was 61.30%and 57.56%,respectively(P=0.045),and the LBR was 44.79%and 36.16%,respectively(P<0.001).For FBT cycles with poor-quality embryo,the CPR at day 5 and 6 was 48.61%and 40.89%,respectively(P=0.006),and the LBR was 31.71%and 25.74%,respectively(P=0.019).The CPR for FBT cycles with good-quality embryo was statistically higher at day 6 than that at day 5 with poor-quality embryo transferred(57.56%vs.48.61%,P=0.001).Maternal age,anti-Müllerian hormone(AMH),endometrial thickness,embryo quality,and the day of blastocyst expansion were independently correlated with the CPR and LBR.The FBT cycles at day 5 had significantly higher CPR(adjusted odds ratio[OR]=1.246,95%confidence intervals[CI]:1.097–1.415,P=0.001)and LBR(adjusted OR=1.435,95%CI:1.258–1.637,P<0.001)than those at day 6.Conclusion The embryo quality is the primary indicator for FBT cycles.Day 5 blastocysts should be preferred when the quality of embryo at day 5 is the same as that at day 6.

Influence of Acupuncture on Idiopathic Male Infertility in Assisted Reproductive Technology

The clinical effects of acupuncture on idio pathic male infertility in sperm parameter and on therapeutic results in assiste d reproductive technology were investigated. 22 patients failed in intracytoplas mic sperm injection (ICSI) with idiopathic male infertility were treated with ac upuncture twice weekly for 8 weeks, followed by ICSI treatment again. The sperm concentration, motility, morphology, fertilization rates and embryo quality were observed. Quick sperm motility after acupuncture(18.3 %±9.6 %) was significa ntly improved as compared with that before treatment (11.0 %±7.5 %, P <0.01) . The normal sperm ratio was increased after acupuncture (21.1 %±10.4 % vs 16.2 %± 8.2 %, P <0.05). The fertilization rates after acupuncture (66.2 %) were obviously higher than that before treatment (40.2 %, P <0.01). There was no significant difference in sperm concentration and general sperm motility between before and after acupuncture. The embryo quality after acupuncture was improved , but the difference between them was not significant ( P >0.05). Acupuncture can improve sperm quality and fertilization rates in assisted reproductive techn ology.

Derivation and characterization of human embryonic stem cell lines from poor quality embryos

Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers on day 3 were cultured in a blastocyst medium for 2 days, and 32 early blastocysts were further cultured in a blastocyst optimum culture medium for additional 2 days so that the inner cell masses （ICMs） could be identified and isolated easily. The ICMs of 17 blastocysts were isolated by a mechanical method, while those of the other 15 blastocysts were isolated by immunosurgery. All isolated ICMs were inoculated onto a feeder layer for subcultivation. The rates of ICM attachment, primary ICM colony formation and the efficiency of hESC derivation were similar between the ICMs isolated by the two methods （P〉0.05）. As a result, four new hESC lines were established. Three cell lines had normal karyotypes and one had an unbalanced Robertsonian translocation. All cell lines showed normal hESC characteristics and had the differentiation ability. In conclusion, we established a stable and effective method for hESC isolation and culture, and it was confirmed that the mechanical isolation was an effective method to isolate ICMs from poor embryos. These results further indicate that hESC lines can be derived from poor quality embryos discarded by IVF laboratories.

Random sperm DNA fragmentation index is not associated with clinical outcomes in day-3 frozen embryo transfer

Damage to sperm DNA was proposed to play an important role in embryonic development.Previous studies focused on outcomes after fresh embryo transfer,whereas this study investigated the influence of sperm DNA fragmentation index(DFI)on laboratory and clinical outcomes after frozen embryo transfer(FET).This retrospective study examined 381 couples using cleavage-stage FET.Sperm used for intracytoplasmic sperm injection(ICSI)or in vitro fertilization(IVF)underwent density gradient centrifugation and swim up processing.Sperm DFI had a negative correlation with sperm motility(r=−0.640,P<0.01),sperm concentration(r=−0.289,P<0.01),and fertilization rate of IVF cycles(r=−0.247,P<0.01).Sperm DFI examined before and after density gradient centrifugation/swim up processing was markedly decreased after processing(17.1%vs 2.4%,P<0.01;65 randomly picked couples).Sperm progressive motility was significantly reduced in high DFI group compared with low DFI group for both IVF and ICSI(IVF:46.9%±12.4%vs 38.5%±12.6%,respectively;ICSI:37.6%±14.1%vs 22.3%±17.8%,respectively;both P<0.01).The fertilization rate was significantly lower in high(≥25%)DFI group compared with low(<25%)DFI group using IVF(73.3%±23.9%vs 53.2%±33.6%,respectively;P<0.01)but was equivalent in high and low DFI groups using ICSI.Embryonic development and clinical outcomes after FET were equivalent for low and high DFI groups using ICSI or IVF.In this study,sperm DFI did not provide sufficient information regarding embryo development or clinical outcomes for infertile couples using FET.

Recent progress in the treatment of women with diminished ovarian reserve

Diminished ovarian reserve(DOR)refers to a decrease in the number and/or quality of oocytes in the ovary,accompanied by a decline in reproductive potential,which is generally related to advanced age or ovarian disease.In in vitro fertilization(IVF)clinical practice,managing patients with DOR remains one of the most challenging tasks.In recent years,increased research on improving ovarian function has provided us with new insights into treating patients with DOR.Many therapeutic options have been proposed to improve the ovarian function of patients with DOR,yet they are not widely utilized in clinical practice because of limited evidence of safety and effectiveness.In this review,we focus on the mechanisms from animal models and clinical trials that have been applied to the treatment of DOR in recent years,intending to improve IVF outcomes in patients with DOR.Furthermore,new insights and perspectives on the molecular and cellular regulation of follicular development and ovarian reserve are emphasized to provide more clues for research on the treatment of DOR.

Sperm-specific protein ACTL7A as a biomarker for fertilization outcomes of assisted reproductive technology

Obtaining high-quality embryos is one of the key factors to improve the clinical pregnancy rate of assisted reproductive technologies(ART).So far,the clinical evaluation of embryo quality depends on embryo morphology.However,the clinical pregnancy rate is still low.Therefore,new indicators are needed to further improve the evaluation of embryo quality.Several studies have shown that the decrease of sperm-specific protein actin-like 7A(ACTL7A)leaded to low fertilization rate,poor embryo development,and even infertility.The aim of this study was to study whether the different expression levels of ACTL7A on sperm can be used as a biomarker for predicting embryo quality.In this study,excluding the factors of severe female infertility,a total of 281 sperm samples were collected to compare the ACTL7A expression levels of sperms with high and low effective embryo rates and analyze the correlation between protein levels and in-vitro fertilization(IVF)laboratory outcomes.Our results indicated that the ACTL7A levels were significantly reduced in sperm samples presenting poor embryo quality.Furthermore,the protein levels showed a significant correlation with fertilization outcomes of ART.ACTL7A has the potential to be a biomarker for predicting success rate of fertilization and effective embryo and the possibility of embryo arrest.In conclusion,sperm-specific protein ACTL7A has a strong correlation with IVF laboratory outcomes and plays important roles in fertilization and embryo development.