Coenzyme Q10 improves the in vitro maturation of oocytes exposed to the intrafollicular environment of patients on fertility treatment.

OBJECTIVE: To evaluate the impact of patient follicular environment with oxidative stress on oocyte quality. METHODS: Patients on fertility treatment with either advanced maternal age or endometriosis were asked to donate follicular fluid collected during ovum pick-up. Follicular fluid (FF) was added (20%, 10% and 5%; %V/V) to in vitro maturation (IVM) medium with mouse oocytes. Following maturation culture, the oocytes were assessed for meiosis reinitiation. In a second setup, coenzyme Q10 was added to culture medium with FF. In addition to assessing meiotic maturation, a subset of oocytes was assessed for spindle structure and chromosome alignment. RESULTS: Supplementation of IVM medium with FF of patients of advanced maternal age (with or without antioxidants) did not have an effect on the maturation capacity of mouse oocytes. However, the addition of FF of individuals with endometriosis (without antioxidants) in the two highest concentrations affected oocyte maturation (61.5% & 57.0% maturation) compared with the lowest concentration (89.2% maturation) (p<0.05). Supplementation of medium with coenzyme Q10 did not improve the maturation rate of oocytes exposed to the FF of individuals with endometriosis (28.5±13.7%) (p<0.05). Nevertheless, preliminary analysis of spindle abnormality and chromosome alignment revealed that oocytes resuming meiosis in the presence of FF of patients with endometriosis displayed aberrant spindle morphology and chromosomal misalignment. CONCLUSION: The follicular environment of patients with endometriosis severely affected oocyte (nuclear) maturation. In vitro maturation in the presence of coenzyme Q10 appears to be a tool for rescuing oocytes exposed to such follicular environment.

Oxidative balance in follicular fluid of ART patients of advanced maternal age and blastocyst formation.

OBJECTIVE: To evaluate the follicular fluid oxidative balance of infertile patients of advanced-maternal-age and the correlation between oxidative imbalance in the follicular fluid and the embryological outcomes. METHODS: Follicular fluid (FF) from infertile patients of advanced-maternal-age undergoing ART treatments were collected and frozen in cryovials at -86°C until further use, and analyzed at the Biochemistry and Nutrition Institute of San Marcos University. As controls, we used FF from oocyte donors. The FF was then assayed for oxidative balance by ABTS, FRAP and TBARS assays. In order to establish the correlation between oxidative balance and embryo quality, we correlated the number of usable blastocysts (freshly transferred or frozen blastocysts) with the results from ABTS, FRAP and TBARS. RESULTS: Follicular fluid from patients of Advanced-maternal-age (AMA group) significantly differed from the values found in the control group; the ABTS value was higher and the FRAP value was lower, in comparison to the FF from oocyte donors (control group). The lipid peroxidation was not different between those two groups. Furthermore, there was no significant correlation among the results of the assays, or when correlated with the proportion of usable blastocysts. CONCLUSION: Ovarian oxidative balance seems to be critical for oocyte quality in advanced-maternal-age patients; however, we still need more studies on oxidative stress indicators, on a larger set of patients.

Occurrence of ovarian follicular dominance during stimulation for IVM impacts usable blastocyst yield.

OBJECTIVE: To evaluate the influence of ovarian follicular dominance on the outcome of oocyte in-vitro maturation. METHODS: This retrospective cohort study included 21 patients with polycystic ovaries or polycystic ovary syndrome (Rotterdam criteria, 2004) subjected to 24 invitro maturation (IVM) cycles between October 2015 and January 2017. Patients undergoing IVM received minimal gonadotropin stimulation starting on day 2 or 3 of the cycle; ovum pick-up typically occurred on days 6 to 8. No hCG-trigger shot was given. Following 30h of IVM, mature oocytes were inseminated by ICSI and the resulting embryos cultured up to the blastocyst stage. RESULTS: Ovarian follicular dominance was observed in nine of the 24 IVM cycles. Oocyte IVM yielded an overall maturation rate of 69.3±23.8%, and no difference was observed when the groups with or without a dominant follicle were assessed independently. The rates of fertilization and usable blastocysts per fertilized oocyte, mature oocyte (Metaphase II) or cumulus-oocyte-complex were nearly three times higher (28.7±22.5%) in the group without ovarian follicular dominance. No differences were found in the clinical pregnancy rates attained by the individuals with or without a dominant follicle after 21 vitrified-warmed blastocyst transfer cycles. CONCLUSION: Occurrence of ovarian follicular dominance during hormonal stimulation for in-vitro maturation negatively impacted embryological outcomes. Strategies devised to limit the appearance of ovarian follicular dominance must be further explored.

Diez años de reproducción asistida de alta complejidad en los Andes del Perú / Ten years of high complexity assisted reproduction techniques in the Peruvian Andes

Introducción: La infertilidad es un diagnóstico cada vez más frecuente, tanto a nivel del mar como en las grandes alturas, y el desarrollo de técnicas de reproducción asistida de alta complejidad en los Andes del Perú es imprescindible. Objetivo: Definir los resultados de estas técnicas en una institución a 3 330 metros sobre el nivel del mar. Diseño: Estudio descriptivo y retrospectivo. Institución: Centro de Fertilidad y Ginecología del Sur, Cusco, Perú. Pacientes: 576 pacientes con infertilidad atendidas entre los años 2005 al 2015. Métodos: Se revisó los registros de los procedimientos realizados y se estableció las tasas de embarazo clínico según la edad y días de cultivo, tanto en ovocitos autólogos como en ovodonación. Lasignificancia estadística de las diferencias encontradas fue analizada por medio de la prueba exacta de Fisher. Principal medida de resultados: Tasa de embarazo clínico. Resultados: Las tasas de embarazo clínico fueron: 30,4% en ovocitos autólogos, 42,4% en ovodonación y 25,0% en maduración in vitro de ovocitos. La frecuencia de embarazo doble fue 31,6%, y triple, 7,9%. Conclusiones: Las tasas de embarazo clínico fueron similares a las observadas internacionalmente. Destaca la necesidad de estrategias para disminuir la pluralidad de los embarazos, así como para mejorar la tasa de embarazo en las mujeres ≥ 37 años.

Introduction: Infertility is currently a more frequent diagnosis, at sea level and at high altitudes, and development of high complexity assisted reproduction techniques in the Andes of Peru is essential. Objective: To define the results of the application of these techniques in an institution at 3 330 meters above sea level. Design: Descriptive and retrospective study. Setting: Centro de Fertilidad y Ginecología del Sur, Cusco, Peru. Patients: 576 patients with infertility attended between years 2005-2015. Methods: Records of the procedures performed were reviewed and rates of clinical pregnancy according to age and days of culture were established for both autologous oocytes and ovodonation. Statistical significance of differences found was analyzed by Fisher exact test. Main outcome measure: Rate of clinical pregnancy. Results: Rates of clinical pregnancy were: 30.4% in autologous oocytes, 42.4% in ovodonation, and 25% in in vitro maturation oocytes. Frequency of twin pregnancy was 31.6%, and of triple pregnancy, 7.9%. Conclusions: Rates of clinical pregnancy were similar to those observed internationally. This emphasizes the need of strategies to diminish the plurality of pregnancies, as well as to improve the rate of pregnancy in women ≥ 37 years old.

Embryotoxicity of serum from women smoking cocaine base paste (CBP).

OBJECTIVE: Using a mouse embryo bioassay, we examined the embryotoxic properties of serum obtained from eight women who had ingested cocaine base paste (CBP) for a period of 1-3 years. STUDY DESIGN: Two-cell stage mouse embryos were cultured in the presence of serum extracted from eight CBP-smoking or control women. After 48h, embryos were evaluated to determine their developmental stage and then processed by Tarkoswki's technique to determine the proportion of embryos with micronuclei, in order to establish the number of micronuclei/embryo. RESULTS: Serum from CBP patients diminished the percentage of embryos progressing to the compacted morula stage, while increasing the proportion of embryos with micronuclei and the corresponding micronuclei/embryo ratio. CONCLUSION: These findings provide the first experimental evidence of embryotoxic compounds in the serum of CBP-smoking women. This study highlights the reproductive risk of chronically ingested CBP and demonstrates the need for further studies.