Mature oocyte dysmorphisms may be associated with progesterone levels, mitochondrial DNA content, and vitality in luteal granulosa cells.

PURPOSE: To identify whether follicular environment parameters are associated with mature oocyte quality, embryological and clinical outcomes. METHODS: This retrospective study examined 303 mature oocytes from 51 infertile women undergoing ICSI cycles between May 2018 and June 2021. Exclusion criteria consisted of advanced maternal age (> 36 years old), premature ovarian failure, obesity in women, or use of frozen gametes. Luteal granulosa cells (LGCs) were analyzed for mitochondrial DNA/genomic (g) DNA ratio and vitality. The relationships between hormone levels in the follicular fluid and oocyte features were assessed. Quantitative morphometric measurements of mature oocytes were assessed, and the association of LGC parameters and oocyte features on live birth rate after single embryo transfer was examined. RESULTS: Results indicated an inverse correlation between the mtDNA/gDNA ratio of LGCs and the size of polar body I (PBI). A 4.0% decrease in PBI size was observed with each one-unit increase in the ratio (p = 0.04). Furthermore, a 1% increase in LGC vitality was linked to a 1.3% decrease in fragmented PBI (p = 0.03), and a 1 ng/mL increase in progesterone levels was associated with a 0.1% rise in oocytes with small inclusions (p = 0.015). Associations were drawn among LGC characteristics, perivitelline space (PVS) debris, cytoplasmic inclusions, PBI integrity, and progesterone levels. Certain dysmorphisms in mature oocytes were associated with embryo morphokinetics; however, live birth rates were not associated with follicular parameters and oocyte quality characteristics. CONCLUSION: Follicular markers may be associated with mature oocyte quality features.

Stress Management during the Intracytoplasmic Sperm Injection Cycle May Slow Down First Embryo Cleavage and Accelerate Embryo Compaction: A Pilot Randomized Controlled Trial.

INTRODUCTION: A firm consensus on the effectiveness of psychological interventions during infertility treatment has not been reached yet in terms of mental health and pregnancy rates. Moreover, the influence of these interventions on embryo cleavage kinetics has not been investigated. OBJECTIVE: The aim of this work was to study whether stress management in couples undergoing an intracytoplasmic sperm injection (ICSI) cycle influences stress levels, mitochondrial DNA (mtDNA) levels in granulosa cells, and cleavage-stage embryos. METHODS: Infertile couples were randomized into a treatment as usual (TAU) group (n = 30) and stress management program (SMP) group (n = 29) at the beginning of an ICSI cycle. Couples in the SMP group attended education and relaxation sessions at each visit to the clinic for folliculometry. The perceived stress scale (PSS) was used to assess stress levels at the beginning and end of the cycle. Moreover, mtDNA levels of granulosa cells and embryo morphokinetics were evaluated. RESULTS: Post-intervention, women in the SMP group had significantly lower PSS scores than their initial PSS (p < 0.001; effect size, ES = 0.5) and than the final PSS of the TAU group (p = 0.02; ES = 0.09). Additionally, mtDNA levels were significantly lower in luteal granulosa cells of the SMP group than the TAU group (p = 0.02). An earlier time of pronuclei appearance (p = 0.03) and time to 2 cells (p = 0.015) and a faster time to full compaction (p = 0.045) were detected in the embryos of the SMP group compared with the TAU group. CONCLUSION(S): The implemented program may reduce stress levels, retard first embryo cleavage, and accelerate embryo compaction. Further studies with an active control group are needed to confirm these results.

Neurophysiology of cognitive behavioural therapy, deep breathing and progressive muscle relaxation used in conjunction with ART treatments: a narrative review.

BACKGROUND: Infertility is defined as the failure to achieve clinical pregnancy after 12 months of regular unprotected intercourse. It could be due to male or female factors, each requiring different treatment options. ART treatment exposes couples to numerous psychological stressors. Therefore, it has been recommended by the ESHRE Psychology and Counselling Guideline Development Group recently that psychosocial support should be offered as a complementary therapy during infertility treatments. In this context, the efficiency of different psychological interventions, such as cognitive behaviour therapy (CBT), deep breathing (DB), and progressive muscle relaxation (PMR), was evaluated in several clinical trials in terms of couples' mental health and pregnancy outcomes. OBJECTIVE AND RATIONALE: The neurophysiology of CBT, DB and PMR, which are used in interventional studies, in both men and women undergoing ART, has not yet been fully elucidated. This review represents a comprehensive report, aiming to collate novel insights into the neurobiological processes and physiological mechanisms that occur during the practice of CBT, DB and PMR. SEARCH METHODS: PubMed, Google Scholar and Cochrane Library were interrogated to conduct this comprehensive literature review. The search was carried out using combinations of MeSH terms and keywords: infertility, assisted reproductive techniques, IVF, ICSI, emotions, psychological stress, cognitive behavioural therapy, mind-body therapies and relaxation. Relevant information related to the mechanism of action of stress management techniques were obtained from original articles and reviews published in English without taking into consideration the time of publication. Moreover, as it was not the major focus of the review, only recent systematic reviews (2015-2019) pinpointing the effects of psychological interventions on infertility treatment outcomes were also retrieved from the above-mentioned databases. OUTCOMES: CBT, DB and PMR may modify the activity of stress-related brain regions such as the prefrontal cortex, amygdala, hypothalamus and hippocampus, as demonstrated by functional MRI and electroencephalogram studies. Furthermore, applying these techniques was associated with mood improvements and a decline in stress biomarkers, and, hypothetically, reducing stress biomarkers attenuates the stress-induced effects on ART outcomes. WIDER IMPLICATIONS: Increasing the knowledge of fertility staff, researchers and physicians regarding the mechanisms of action of these stress management techniques has several advantages. For instance, understanding the underlying neurophysiological pathways would assist practitioners to engage ART couples in the practice of these techniques. Also, it may enhance the quality of the support programmes and psychological research. Accordingly, this will ensure that these interventions reach their full potential and therefore improve clinical outcomes.

Optimization of The Cell Aggregates Method for Isolation and Purification of Human Granulosa Cells from Follicular Fluid.

BACKGROUND: Aspirated ovarian follicular fluids (FF) contain luteal granulosa cells (LGCs) and other contaminating cell types. Several strategies, such as the antibody binding methods, the flask method, the cell strainer and positive selection of granulosa aggregates after density gradient (DG) centrifugation, were tested as LGC purification methods. Each of these strategies has its own advantages and disadvantages. Positive selection of granulosa aggregates after DG centrifugation is simple, rapid and efficient in terms of LGC recovery. However, it results in a low purity. Here, we aimed to test whether modifying the traditional protocol by collecting the aggregates from the FF, before the DG centrifugation could decrease the percentage of contaminating cells. MATERIALS AND METHODS: In the present prospective study, 32 FF, from 32 women,were randomly assigned into one of the two purification techniques: positive selection of granulosa aggregates from the FF, after DG centrifugation (DG/ Agg, n=16) or positive selection of granulosa aggregates from the FF, before DG centrifugation (Agg/DG, n=16). At the end of each procedure cell count, vitality, morphology and purity of the cell suspension were evaluated. RESULTS: No significant difference was detected in the total number of GCs between DG/Agg and Agg/DG (P>0.05). However, higher percentage of GCs with normal morphology was detected in Agg/DG compared to DG/Agg (P<0.001). Moreover, lower percentages of white blood cells (P<0.01), red blood cells (P<0.001) and epithelial cells (P<0.01) were identified in Agg/DG compared to DG/Agg. CONCLUSION: Here we showed that positive selection of granulosa aggregates from the FF prior to DG technique had a higher purity compared to the traditional protocol. Thus, it could be a method of choice to prepare GCs for research purposes in clinical in vitro fertilization settings.