Morphological, ultrastructural, and molecular aspects of in vitro mouse embryo implantation on human endometrial mesenchymal stromal cells in the presence of steroid hormones as an implantation model

Objective: This experimental study aimed to evaluate the effects of 17beta-estradiol [E2] and progesterone [P4] on the interaction between mouse embryo and human endometrial mesenchymal stromal cells, and gene expressions related to implantation [alphaV and beta3 integrins, interleukin-1 receptor [IL-1R], and leukemia inhibitory factor receptor [LIFR]] using an in vitro twodimensional model

Materials and Methods: In this experimental study, the endometrial stromal cells were isolated enzymatically and mechanically, and cultured to the fourth passage. Next, their immunophenotype was confirmed by flow cytometric analysis as mesenchymal stromal cells. The cells were cultured as either the experimental group in the presence of E2 [0.3 nmol] and P4 [63.5 nmol] or control group without any hormone treatment. Mouse blastocysts were co-cultured with endometrial mesenchymal stromal cells in both groups for 48 hours. Their interaction was assessed under an inverted microscope and scanning electron microscopy [SEM]. Expressions of alphaV and beta3 integrins, LIFR, and IL-1R genes were analyzed by real-time reverse transcription-polymerase chain reaction [RT-PCR]

Results: Similar observations were seen in both groups by light microscopy and SEM. We observed the presence of pinopode-like structures and cell secretions on the apical surfaces of endometrial mesenchymal stromal cells in both groups. The trophoblastic cells expanded and interacted with the mesenchymal monolayer cells. At the molecular level, expression of IL-1R significantly increased in the hormonal treated group compared to the control [P

Conclusion: This study has shown that co-culture of endometrial mesenchymal stromal cells with mouse embryo in media that contained E2 [0.3 nmol] and P4 [63.5 nmol] could effectively increase the expression of IL-1R, which is involved in embryo implantation. However, there were no significant effects on expressions of alphaV and beta3 integrins, LIFR, and on the morphology and ultrastructure of endometrial mesenchymal stromal cells

Saccharin consumption increases sperm DNA fragmentation and apoptosis in mice

Saccharin is an artificial non-caloric sweetener that used to sweeten products such as drinks, candies, medicines, and toothpaste, but our bodies cannot metabolize it. Sodium saccharin is considered as an important factor in tumor promotion in male rats but not in humans. The objective of this study was to investigate the effect of saccharin consumption on sperm parameters and apoptosis in adult mice. Totally 14 adult male mice were divided into 2 groups. Group 1 served as control fed on basal diet and group 2 or experimental animals received distilled water containing saccharin [0.2% w/v] for 35 days. After that, the left cauda epididymis of each mouse was cut and placed in Ham's F10. Swimmed-out spermatozoa were used to analyze count, motility, morphology [Pap-staining] and viability [eosin-Y staining]. Sperm DNA integrity, as an indicator of apoptosis, was assessed by SCD [sperm chromatin dispersion] and terminal deoxynucleotidyl transferase [TUNEL] assay. Following saccharin consumption, we had a reduction in sperm motility with respect to control animals [p=0.000]. In addition, the sperm count diminished [17.70 +/- 1.11 in controls vs. 12.80 +/- 2.79 in case group, p=0.003] and the rate of sperm normal morphology decreased from 77.00 +/- 6.40 in control animals into 63.85 +/- 6.81 in saccharin-treated mice [p=0.001]. Also, we saw a statistically significant increase in rates of sperm DNA damage and apoptosis in experimental group when compared to control one [p=0.001, p=0.002 respectively]. Saccharin consumption may have negative effects on sperm parameters, and increases the rate of sperm DNA fragmentation and apoptosis in mice

Does women's age influence zona pellucida birefringence of metaphase II oocytes in in-vitro maturation program?

In vitro maturation [IVM] is a promising treatment option for certain infertile women. Nowadays, with the aid of PolScope, it has become possible to evaluate zona pellucida [ZP] characteristics as a parameter of oocyte quality. Moreover, quality of oocytes can be influenced by many factors, such as patient's age. The PolScope system is a non-invasive technique to assess birefringent structures such as the meiotic spindle and ZP in living oocytes. The aim was to determine the influence of the woman's age on ZP birefringence, a sign of oocyte quality, and morphology of in-vitro matured human oocytes using non-invasive polarized light [PolScope] microscopy. ZP birefringence and morphology were determined in 105 retrieved oocytes from 58 women undergoing ICSI in two age groups [>/= 30 years and <30 years]. The immature oocytes were selected and after IVM, the quality of metaphase II [MII] oocytes was assessed. The oocytes abnormalities were classified as intracytoplasmic and extracytoplasmic abnormalities. Oocyte maturation rates were significantly reduced in >/= 30 year's women [56%] in comparison with other age group [80.7%]. In addition, the ZP birefringence was significantly higher in MII oocytes in the younger group compared with the older group [76.2% vs.38.1%; p=0.00]. Following morphologic assessment, the rates of oocytes with extracytoplasmic [p=0.02] and both abnormalities [extra- and intracytoplasmic] [p=0.01] were higher in aged versus the younger women. There was a positive relationship between advanced maternal age with decreased ZP birefringence and oocyte morphological quality in in-vitro matured human oocytes