Effect of Administration of Single Dose GnRH Agonist in Luteal Phase on Implantation Rate of Fresh ICSI Cycles

The luteal phase (LP) in the fresh ICSI cycle is insufficient, adequate LP support is one of the approved treatments for improving implantation and pregnancy rates. It is generally known that the LP is inadequate after ovarian stimulation due to negative from supra-physiological blood levels of steroids released by numerous corporal luteal, LH concentrations are low during the luteal phase. In this study, patients were divided into two groups: (40) patients as study group; those who received GnRHa (Decapeptil 0.1 mg), three days after embryo transfer, in addition to conventional luteal phase support (LPS) in the LP to increase the implantation and pregnancy rate in IVF; and their control group (40) received standard LPS only. On the second day of stimulation, blood samples for FSH, LH, TSH, E2, and prolactin were taken. On the day of ovulation induction, measure E2, progesterone, and LH; and on the day of embryo transfers, measure progesterone and LH. The overall characteristics of the patients in both groups were not significantly different. There was also no significant change in the number of total oocytes, mean of metaphase II oocytes percent, cleavage rate, grade I embryo percent, or serum hormones level between the study and control groups (p > 0.05). GnRH agonist treatment in the luteal phase improves clinical pregnancy and implantation rate in fresh ICSI cycles but is not statistically significant.

Association Between Urine and Serum Estradiol Levels in In Vitro Fertilization Cycles

Objectives To study the correlation between urine and serum estradiol (E2) controlled ovarian hyperstimulation (COH) Methods This is a cross-sectional analytical study that was conducted in a tertiary care hospital. Seventy-seven urine and blood samplings were collected from infertile women who were treated with COH. An electrochemiluminescent immunoassay was performed to evaluate E2 levels between urine and serum samples on the 6th day and the day of ovarian trigger. In addition, the correlations were evaluated between urine E2 level and number of follicles, retrieved, metaphase II oocytes, and fertilization rate. A sub-analysis was performed for age, responding status and BMI. Results Seventy-seven infertile women were recruited. The medians of serum and urine E2 level (E2/creatinine) levels on the day 6th of ovarian stimulation were 833.20 pg/ml (IQR; 516.90–1,371.00) and 3.67 (IQR; 2.84–4.81), respectively. On the day of ovarian trigger, the median of serum E2 level was 2,113.00 pg/ml (IQR; 1,382.00–3,885.00) and urine E2 level (E2/creatinine) was 6.84 (IQR; 5.34–8.70). The correlation between serum and urine E2 level on day 6th was 0.53 and the day of ovarian trigger was 0.59, p < 0.001. Moreover, the correlations of urine E2 level on the day of ovarian trigger to number of follicles, number of oocytes retrieved, metaphase II oocytes and fertilization rate were 0.57, 0.58, 0.61, and 0.64 (P < 0.001). Conclusion The urine E2 level was moderately correlated to serum E2, number of follicles growth, oocytes retrieved and also fertilization rate.

P–342 Are blastulation and clinical pregnancy rates in women with endometriomas different than those without?

Study question Does the presence of endometrioma during ovarian stimulation affect blastulation and clinical pregnancy rates (CPR)? Summary answer Blastulation rates were similar in women with endometrioma compared to women without. Likewise, CPR were comparable. What is known already Although relationship of endometriosis and subfertility is well-established, its mechanism is still under investigation. Decreased oocyte quality, resulting from anatomical and/or inflammatory factors is one of the prominent culprits. Most studies regarding endometriosis and oocyte quality are highly heterogeneous and effect of endometriosis on oocyte quality is yet to be determined. Blastulation is thought as a surrogate marker for oocyte quality. Thus, it may be possible that detrimental effect of the presence of endometrioma during ovarian stimulation can be indirectly assessed by blastulation. Study design, size, duration Records of all women who underwent assisted reproductive technology treatment at Koc University Hospital Assisted Reproduction Unit between 2016 and October 2020 were screened for this retrospective study. All women who had endometrioma(s) during ovarian stimulation were included in the study group (EG) (n = 71). They were matched with women diagnosed with tubal factor or unexplained infertility who underwent oocyte pickup within the same period to form the control group (CG) (n = 104). Participants/materials, setting, methods All women underwent antagonist or long protocol. All embryos were cultured until blastocyst stage regardless of the number of oocytes or embryos available. Size/location of endometriomas, number of oocytes retrieved, number of available blastocysts, positive pregnancy test per cycle and clinical pregnancy rate per cycle were recorded. Blastulation rate was calculated as number of available blasts divided by the number of metaphase-II oocytes. Embryos were transferred in a fresh or artificially prepared frozen-thawed cycle. Main results and the role of chance There were 71 women in EG and 104 women in CG, which included 30 women with tubal and 74 with unexplained infertility. Median endometrioma size was 26 mm(22–33). Twenty-three patients in EG had history of endometrioma excision (31.3%). Median age [35.0 years (31.0–39.0) vs 34 (32.0–36.0), p = 0.26] and serum AMH levels [1.8 (1.1 - 4.2) vs 2.3 (1.3 - 3.7) ng/dL, p = 0.91] were similar in EG and CG, respectively. Body mass index in kg/m2 [21.8 (20.2–24.6) vs 24 (21.5–27.9), p &lt; 0.01] and infertility duration in years [2 (1–2.6) vs 3 (2–5), p &lt; 0.01] were significantly lower in EG. Number of retrieved oocytes [8 (5–12) vs 12 (7–15.8), p &lt; 0.01)] and metaphase-II oocytes [6 (4–10) vs 8.5 (6–12), p &lt; 0.01] were lower in EG group compared to CG group. However, blastulation rate per MII oocyte were similar between the EG and CG [(0.25 (0.20–0.41) vs 0.30 (0.14–0.50), respectively, p = 0.58]. Adjusted analysis for age and number of MII oocytes revealed similar finding. Positive pregnancy test per cycle was similar at 53.5% vs 61.5% in EG and CG, respectively (p = 0.3). CPR were similar between the EG and CG (45% vs 58%, respectively, p = 0.10). Limitations, reasons for caution Retrospective design, lack of live birth information are the main limitations of our study. Wider implications of the findings: Presence of endometrioma during ovarian stimulation does not seem to adversely affect blastulation rates. While this is reassuring regarding oocyte quality, further research is required to assess its effect on live birth. Trial registration number Not applicable

P–596 Association of oleic acid production in cumulus-granulosa cells with glutathione of in vitro matured oocytes

Study question Does oleic acid production in cumulus-granulosa cells affect glutathione levels of in vitro matured oocytes? Summary answer Oleic acid availability in cumulus-granulosa cells is associated with a higher glutathione level in in vitro matured oocytes. What is known already The monounsaturated fatty acid oleic acid is de novo synthesized by desaturation of stearic acid and can promote steroidogenesis and oocyte development in vitro. The endogenous antioxidant glutathione content in metaphase II oocyte is significantly higher than immature stages and is related to the normal oocyte maturation. Study design, size, duration Mouse germinal vesicles were co-cultured for 24 hours, during in vitro maturation, with granulosa cells treated with a specific inhibitor of oleic acid synthesis alone or in combination with oleic acid. Participants/materials, setting, methods Fluorescence staining was used to assess the glutathione content of mouse metaphase II oocytes following in vitro maturation as an indicator of cytoplasmic maturation. Glutathione was stained using Cell Tracker Blue –CMAC for 30 min at 37 °C. After being washed in fresh media, stained oocytes were photographed by a fluorescence microscope. Cell area and associated fluorescence were quantified in 20 metaphase II mouse oocytes randomly chosen from in vitro matured oocytes for each condition. Main results and the role of chance The intracellular glutathione content was profoundly lower in metaphase II oocytes obtained from co-cultures with inhibitor-treated cumulus-granulosa cells than with the control cumulus cells (–50%, p &lt; 0.01). Oleic acid effectively recovered the negative effect of inhibitor on glutathione level nearly up to the level of the mock-treated cells. Limitations, reasons for caution The findings are limited to metaphase II. Measurement at more advanced stages of oocyte development is of interest. Inhibition of cellular fatty acid synthesis was performed solely with a specific chemical. Wider implications of the findings: Involvement of the oleic acid availability for cumulus-granulosa cells in normal oocyte maturation may be of relevance in reproductive disorders, particularly in the pathological mechanism of impaired oogenesis. Trial registration number 400/3226

Developmental potency of goat embryos produced by intra cytoplasmic sperm injection and in vitro fertilization.

The objective of this study was to standardize novel intracytoplasmic sperm injection (ICSI) technique, production of goat embryos by ICSI, in vitro fertilization (IVF) and comparison of their developmental potency. Immobilized sperm from epididymal semen with breaking tail were injected to mature metaphase ii oocytes by sharp micropipette of 5 µm diameter, as well as in vitro fertilization was carried out. Icsi and IVF derived zygotes were cultured in rvcl-blast-bsa media after chemical activation with ionomycin, chx and 6-dmap. Sham injection was used as a control group. Results suggest that immobilized sperm with breaking tail microinjected by sharp injection pipette after chemical activation is an efficient approach for goat embryo production by ICSI. Satisfactory survival rate (83.2%) was found by using sharp injection pipette of 5 µm diameter. Chemical activation with ionomycin, chx and 6-dmap was found effective with activation rate of 63.33%. Developmental rate of 2 cell, 4–8 cell, 8–16 cell, morula, blastocyst in ICSI and IVF group was 74.44%, 63.33%, 44.44%, 23.33%, 14.44% and 65.16%, 57.42%, 49.03%, 38.71%, 23.23%, respectively. Upon comparison, significantly higher (p < 0.05) percentage of cleavage was found in ICSI group than IVF (73.40 ± 3.12% vs. 64.06 ± 2.44%, respectively). However, morula and blastocyst percentage was significantly higher (p < 0.05) in IVF group than ICSI (23.20 ± 2.73% vs. 37.90 ± 3.15% and 14.56 ± 1.75% vs. 21.05 ± 2.09%). It can be concluded that ICSI with sharp micropipette of 5 µm diameter can be applied to assisted reproduction for in vitro goat embryo production.

Effect of oocyte vitrification on glucose transport in mouse metaphase II oocytes

Oocyte vitrification has significantly improved the survival rate and become the mainstream method for cryopreserving oocytes. Previous studies have demonstrated that the ultrastructure, mitochondrial function, DNA methylation, and histone modification exhibit an irreversible effect after oocyte vitrification. However, little is known about the effects of oocyte vitrification on glucose transport and metabolism. This study aims to determine whether mouse oocyte vitrification causes abnormal glucose metabolism and identify a strategy to correct abnormal glucose metabolism. Furthermore, this study further investigates the effects of oocyte vitrification on glucose uptake, and glucose metabolism, and energy levels. The results indicated that vitrification significantly reduced the glucose transport activity, NADPH, glutathione, and ATP levels, and increased reactive oxygen species levels in oocytes (P < 0.01). Vitrification also reduced the expression of glucose transporter isoform 1 (GLUT1) (P < 0.01). Adding a GLUT1 inhibitor reduced the glucose uptake capacity of oocytes. Furthermore, the inclusion of vitamin C into thawing and culture solutions restored abnormal glucose transportation and metabolism and improved the survival, two-cell embryo, and blastocyst rates of the vitrified groups via parthenogenesis (P < 0.05). Overall, this method may improve the quality and efficiency of oocyte vitrification.