Effects of lactate, super-GDF9, and low oxygen tension during bi-phasic in vitro maturation on the bioenergetic profiles of mouse cumulus-oocyte complex†.

In vitro maturation (IVM) is an alternative assisted reproductive technology with reduced hormone-related side effects and treatment burden compared to conventional IVF. Capacitation (CAPA)-IVM is a bi-phasic IVM system with improved clinical outcomes compared to standard monophasic IVM. Yet, CAPA-IVM efficiency compared to conventional IVF is still suboptimal in terms of producing utilizable blastocysts. Previously, we have shown that CAPA-IVM leads to a precocious increase in cumulus cell (CC) glycolytic activity during cytoplasmic maturation. In the current study, considering the fundamental importance of CCs for oocyte maturation and cumulus-oocyte complex (COC) microenvironment, we further analyzed the bioenergetic profiles of maturing CAPA-IVM COCs. Through a multi-step approach, we (i) explored mitochondrial function of the in vivo and CAPA-IVM matured COCs through real-time metabolic analysis with Seahorse analyzer, and to improve COC metabolism (ii) supplemented the culture media with lactate and/or super-GDF9 (an engineered form of growth differentiation factor 9) and (iii) reduced culture oxygen tension. Our results indicated that the pre-IVM step is delicate and prone to culture-related disruptions. Lactate and/or super-GDF9 supplementations failed to eliminate pre-IVM-induced stress on COC glucose metabolism and mitochondrial respiration. However, when performing pre-IVM culture under 5% oxygen tension, CAPA-IVM COCs showed similar bioenergetic profiles compared to in vivo matured counterparts. This is the first study providing real-time metabolic analysis of the COCs from a bi-phasic IVM system. The currently used analytical approach provides the quantitative measures and the rational basis to further improve IVM culture requirements.

Biochanin-A attenuates DHEA-induced polycystic ovary syndrome via upregulation of GDF9 and BMP15 signaling in vivo.

AIMS: Phytoestrogens can act as natural estrogens owing to their structural similarity to human estrogens. Biochanin-A (BCA) is a well-studied phytoestrogen with a wide variety of pharmacological activities, whereas not reported in the most frequently encountered endocrinopathy called polycystic ovary syndrome (PCOS) in women. PURPOSE: This study aimed to investigate the therapeutic effect of BCA on dehydroepiandrosterone (DHEA) induced PCOS in mice. MAIN METHODS: Thirty-six female C57BL6/J mice were divided into six groups: sesame oil, DHEA-induced PCOS, DHEA + BCA (10 mg/kg/day), DHEA + BCA (20 mg/kg/day), DHEA + BCA (40 mg/kg/day), and metformin (50 mg/kg/day). KEY FINDINGS: The results showed a decrease in obesity, elevated lipid parameters, restoration of hormonal imbalances (testosterone, progesterone, estradiol, adiponectin, insulin, luteinizing hormone, and follicle-stimulating hormone), estrus irregular cyclicity, and pathological changes in the ovary, fat pad, and liver. SIGNIFICANCE: In conclusion, BCA supplementation inhibited the over secretion of inflammatory cytokines (TNF-α, IL-6, and IL-1ß) and upregulated TGFß superfamily markers such as GDF9, BMP15, TGFßR1, and BMPR2 in the ovarian milieu of PCOS mice. Furthermore, BCA reversed insulin resistance by increasing circulating adiponectin levels through a negative correlation with insulin levels. Our results indicate that BCA attenuated DHEA-induced PCOS ovarian derangements, which could be mediated by the TGFß superfamily signaling pathway via GDF9 and BMP15 and associated receptors as first evidenced in this study.

Electroacupuncture Enhances Number of Mature Oocytes and Fertility Rates for In Vitro Fertilization.

Objective: The increasing prevalence of infertility every year is in line with the increasing need for in vitro fertilization (IVF) programs. Failure of oocyte maturation is an obstacle that often causes low success in IVF. According to several studies, controlled ovarian stimulation (COS) has been reported to increase the risk of granulosa-cell apoptosis associated with inhibition of oocyte maturation. This study was conducted to determine the effect of electroacupuncture (EA) on oocyte maturation; rate of fertilization; granulosa-cell apoptosis; and levels of growth-differentiation factor 9 (GDF9) and bone-morphogenetic protein 15 (BMP15) an IVF program. Materials and Methods: A randomized controlled trial was conducted with 24 subjects who were in the IVF program. Subjects were randomly allocated into verum-EA (n = 12) and sham-EA groups (n = 12). Microscopic assessment of oocyte maturation and rate of fertilization was performed by embryologists, and examinations of granulosa-cell apoptosis index (Bax/Bcl-2 ratio), GDF9, and BMP15 were performed, using real-time quantitative polymerase chain reaction and messenger-RNA techniques. Results: There were significant differences in oocyte maturation (P = 0.02) and fertilization rates (P = 0.03) between the verum-EA and sham-EA groups. There were differences in granulosa-cell apoptosis index between the verum-EA and sham-EA groups (P < 0.001). There were significant differences in Bax-protein expression (P = 0.04) and Bcl-2 (P = 0.03) granulosa cells between the verum-EA and sham-EA groups. There was no significant difference in GDF9 levels (P = 0.34) and BMP15 (p = 0.47) between the verum-EA and sham-EA groups. Conclusions: EA can enhance oocyte maturation and fertilization rate, and reduce the granulosa-cell apoptosis index in an IVF program.