D. Castello

Day-3 embryo metabolomics in the spent culture media is altered in obese women undergoing in vitro fertilization

OBJECTIVE To determine whether the global metabolomic profile of the spent culture media (SCM) of day-3 embryos is different in obese and normoweight women undergoing in vitro fertilization (IVF). DESIGN Prospective cohort analysis. SETTING IVF clinic. PATIENT(S) Twenty-eight young, nonsmoking women with normoweight, nonsmoking male partners with mild/normal sperm factors undergoing a first IVF attempt for idiopathic infertility, tubal factor infertility, or failed ovulation induction: obese ovulatory women (n = 12); obese women with polycystic ovary syndrome (PCOS; n = 4); normoweight ovulatory women (n = 12). INTERVENTION(S) Fifty μl of SCM collected from two day-3 embryos of each cohort. MAIN OUTCOME MEASURE(S) Metabolomic profiling via ultrahigh performance liquid chromatography coupled to mass spectrometry of SCM from a total of 56 embryos. RESULT(S) The untargeted metabolomic profile was different in obese and normoweight women. Partial least squares discriminant analysis resulted in a clear separation of samples when a total of 551 differential metabolites were considered. A prediction model was generated using the most consistent metabolites. Most of the metabolites identified were saturated fatty acids, which were detected in lower concentrations in the SCM of embryos from obese women. The metabolomic profile was similar in obese women with or without PCOS. CONCLUSION(S) The metabolomic profile in the SCM of day-3 embryos is different in normoweight and obese women. Saturated fatty acids seem to be reduced when embryos from obese patients are present. CLINICAL TRIAL REGISTRATION NUMBER NCT01448863.

Outcome of cryotransfer of embryos developed from vitrified oocytes: double vitrification has no impact on delivery rates

OBJECTIVE Evaluate the outcome of cryotransfer of embryos developed from vitrified oocytes. DESIGN Retrospective cohort study. SETTING Private university-affiliated IVF center. PATIENT(S) Women undergoing warming cycles in which vitrified embryos were developed from vitrified or fresh oocytes. INTERVENTION(S) Vitrification by the Cryotop open device. MAIN OUTCOME MEASURE(S) Delivery rate (DR) per warming cycle. RESULT(S) A total of 471 warming cycles of 796 vitrified embryos developed from vitrified oocytes (group 1) and 2,629 warming cycles of 4,394 vitrified embryos derived from fresh oocytes (group 2) were evaluated. Overall survival rates were 97.2% [95% confidence interval [CI] 95.9%-98.6%] vs. 95.7% [95% CI 94.9-96.4], respectively. DRs per warming cycle were 33.8% (group 1) and 30.9% (group 2). Double vitrification had no effect on DR (odds ratio [OR] 0.877, 95% CI 0.712-1.080). Confounding factors (ovum donation or autologous cycles; day-3 or blastocyst embryo transfer [ET]; natural or hormonal replacement therapy for ET; single or double ET; previous cycles, number of oocytes, doses of gonadotropins and E2 levels on the day of hCG) did not modify the effect of double vitrification on DR (OR 0.872, 95% CI 0.702-1.084). CONCLUSION(S) Vitrification at early cleavage or blastocyst stage of embryos obtained from previously vitrified oocytes has no effect on DR/warming cycle.

Improvement in reproductive outcome through artificial oocyte activation on fertilization failure cases: a cohort study

MATERIALS AND METHODS: We described the outcome from 273 oocytes from 66 patients who underwent first attempt of ICSI without AOA, TABLE 1. Outcomes by insemination method ICSI IVF P-value High quality total blastulation ratey Mean (SD) (n[702 cycles) 28% (28) (n[381 cycles) 37% (32) <0.05 High quality total blastulation rate -Split groupy Mean (SD) (n[94 cycles) 43% (29) (n[94 cycles) 49% (31) <0.05 First cycles only ICSI IVF P-value High quality total blastulation rate y Mean (SD) (n[365 cycles) 32% (29) (n[251 cycles ) 43% (32) <0.05 High quality total blastulation rate Split-group y Mean (SD) (n[78 cycles) 44% (32) (n[78 cycles) 50% (33) <0.05 y(# day 5 High quality blast + # day 6 High quality blast)y (# 2pn # trans on day 3) FERTILITY & STERILITY_ e221 getting either fertilization failure or low percentage of fertilization (<30%) and were compared with 620 oocytes from the same cohort of patients (84 cycles) in which a new attempt (48 with one cycle and 18 patients with two cycles) were performed with AOA. Study period included between April 2013 and September 2016. The injection of the oocytes by AOAwas carried out by injecting the spermatozoa together with a previous phase of buffered media with Ica (1/3 of pipette in 40x magnification microscope). Later, they were kept for ten minutes in the incubator with fertilization media and ICa in a 37 _ C, 6%CO2 atmosphere. Embryo culture was carried out in standard incubator under culture conditions of 37_C, 6% CO2, 5% O2 atmosphere. Fertilization, pregnancy, implantation and abortion rates were analysed and compared in both groups by X2, t-Student and ANOVA tests when were needed.

Pre-clinical validation of a closed surface system (Cryotop SC) for the vitrification of oocytes and embryos in the mouse model

Vitrification is currently a well-established technique for the cryopreservation of oocytes and embryos. It can be achieved either by direct (open systems) or indirect (closed systems) contact with liquid nitrogen. While there is not a direct evidence of disease transmission by transferred cryopreserved embryos, it was experimentally demonstrated that cross-contamination between liquid nitrogen and embryos may occur, and thus, the use of closed devices has been recommended to avoid the risk of contamination. Unfortunately, closed systems may result in lower cooling rates compared to open systems, due to the thermal insulation of the samples, which may cause ice crystal formation resulting in impaired results. In our study, we aimed to validate a newly developed vitrification device (Cryotop SC) that has been specifically designed for being used as a closed system. The cooling and warming rates calculated for the closed system were 5.254 °C/min and 43.522 °C/min, respectively. Results obtained with the closed system were equivalent to those with the classic Cryotop (open system), with survival rates in oocytes close to 100%. Similarly, the potential of the survived oocytes to develop up to good quality blastocysts after parthenogenetic activation between both groups was statistically equivalent. Assessment of the meiotic spindle and chromosome distribution by fluorescence microscopy in vitrified oocytes showed alike morphologies between the open and closed system. No differences were found either between the both systems in terms of survival rates of one-cell stage embryos or blastocysts, as well as, in the potential of the vitrified/warmed blastocysts to develop to full-term after transferred to surrogate females.