Endometrial Mesenchymal Stromal Cells (endMSCs) are multipotent cells with immunomodulatory and pro-regenerative activity which is mainly mediated by a paracrine effect. The exosomes released by MSCs have become a promising therapeutic tool for the treatment of immune-mediated diseases. More specifically, extracellular vesicles derived from endMSCs (EV-endMSCs) have demonstrated a cardioprotective effect through the release of anti-apoptotic and pro-angiogenic factors. Here we hypothesize that EV-endMSCs may be used as a co-adjuvant to improve in vitro fertilization outcomes and embryo quality. Firstly, endMSCs and EV-endMSCs were isolated and phenotypically characterized for in vitro assays. Then, in vitro studies were performed on murine embryos co-cultured with EV-endMSCs at different concentrations. Our results firstly demonstrated a significant increase on the total blastomere count of expanded murine blastocysts. Moreover, EV-endMSCs triggered the release of pro-angiogenic molecules from embryos demonstrating an EV-endMSCs concentration-dependent increase of VEGF and PDGF-AA. The release of VEGF and PDGF-AA by the embryos may indicate that the beneficial effect of EV-endMSCs could be mediating not only an increase in the blastocyst’s total cell number, but also may promote endometrial angiogenesis, vascularization, differentiation and tissue remodeling. In summary, these results could be relevant for assisted reproduction being the first report describing the beneficial effect of human EV-endMSCs on embryo development.

The supplementation of embryo culture medium with extracellular vesicles from endometrialderived mesenchymal stem cells increases the quality of aged embryos

Introduction: With the health professional's better alertness to the benefits of breastfeeding, the rates of discontinuation show some discrete signs of improvement. Objective: To determine the prevalence of breastfeeding during the first six months of life within the pediatric population served by the Barão do Corvo Primary Care Center, and analyze its relation with age, parity, social-economic backgrounds, maternal profession and education, experience in breastfeeding, level of information and advice and health professional's practices. Methods: An inquiry was applied to the mothers of infants from 6 to12 months old brought to that primary care unit during the month of March 2003. Results: We obtained 91 questionnaires (15% of the target population). On the day of discharge from maternity 98% of mothers were breastfeeding; 56% in the first month of life (the higher rate of weaning) and 34% in the sixth month. The frequency of the reasons why breastfeeding was interrupted was determined. The introduction of formula was indicated by the pediatrician in 39% of the cases. Factors like first time mother, adolescent mother, no experience in breastfeeding, lower education, and having received bottle supplementation in the maternity, relates negatively with the duration of breastfeeding. Conclusion: The prevalence of breastfeeding at birth is excellent, but at six months is far away from OMS goals, of 50% babies' exclusively breastfed by this age. The identification of risk factors for early termination of breastfeeding allows the implementation of public health interventions aimed at improving the prevalence of exclusive breastfeeding for at least the first 4 to 6 months of babies' life.

Advanced age reduces the success of in vitro fertilization (IVF) being this effect partly mediated by an overproduction of reactive oxygen species (ROS) that trigger apoptosis. It has been demonstrated that extracellular vesicles derived from endometrial mesenchymal stem cells (EV‐endMSCs) exert an antioxidant effect and can be used as IVF coadjutants. In this work, endMSCs were isolated from human menstrual blood (n = 4) and characterized according to multipotentiality and surface marker expression prior EV‐endMSCs isolation. Oocytes were obtained from 21 B6D2 mice (24 weeks) and coincubated with sperm from young males (8–12 weeks). Presumptive zygotes were incubated in the presence of 0, 10, 20, 40 or 80 μg/ml of EV‐endMSCs in KSOM medium. Blastocyst yield was evaluated, and 25 blastocysts per group were used for qPCR. Blastocyst rate was 29.4% in control; 45.2% for 10 μg/ml, 62.9% for 20 μg/ml, 55.5% for 40 μg/ml and 53.8% in the 80 μg/ml (n = 124–130 oocytes) being all the increases significantly different when compared against control (p < 0.05). The 20–80 μg/ml treatments decreased the expression of glutathione peroxidase (Gpx1), and the 10–40 μg/ml treatments reduced the expression of superoxide dismutase (Sod1; p < 0.05) compared to control; Bax mRNA expression did not vary. Our results suggest that the increased developmental competence of the embryos could be partly mediated by the EV‐endMSCs’ ROS scavenger activity.

BackgroundVitrification is the safest method to cryopreserve oocytes, however the process alters mitochondrial function resulting from increased reactive oxygen species (ROS) production. Our aim was to alleviate ROS stress in vitrified mice oocytes using N-acetylcysteine (NAC at 1 mM), to improve the oocyte’s developmental competence.ResultsHence, four experimental groups were compared: fresh oocytes (F-C), vitrified oocytes (V-C), NAC addition prior to oocyte vitrification (V-NAC-Pre) and NAC addition after vitrification (V-NAC-Post). V-NAC-Pre and V-NAC-Post exhibited higher levels of mitochondrial polarization compared to vitrified oocytes (36.5 ± 3.1, 37.7 ± 1.3 and 27.2 ± 2.4 measured as the spatial coefficient of variation/oocyte respectively, mean ± SEM; p < 0.05). However, ROS production increased in vitrified oocytes added with NAC compared to the vitrified control (1124.7 ± 102.1 [V-NAC-Pre] and 1063.2 ± 82.1 [V-NAC-Post] vs. 794.6 ± 164.9 [V-C]; arbitrary fluorescence units/oocyte, mean ± SEM; p < 0.05). ATP significantly decreased in V-NAC-Pre compared to V-NAC-Post oocytes (18.5 ± 6.9 vs. 54.2 ± 4.6 fmol/oocyte respectively, mean ± SEM; p < 0.05), and no differences were observed between V-NAC-Post, F-C and V-C groups. Blastocyst rates derived from F-C oocytes was higher than those derived from V-NAC-Pre (90.7 ± 1.8 vs. 79.1 ± 1.8, respectively, mean % ± SEM,; p < 0.05) but similar to those derived from V-NAC-Post (90.7 ± 1.8, mean % ± SEM, p > 0.05). Total blastomere count of blastocysts derived from V-NAC-Post after in vitro fertilization (IVF) was higher than embryos produced from V-C.ConclusionsThe addition of NAC after vitrification improves the quality of vitrified mature murine oocytes while its addition prior to vitrification is advised against.