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INTRODUCCIÓN: La infertilidad es definida por la Organización Mundial de la Salud (OMS) como la patología del sistema reproductor caracterizada por la incapacidad de lograr un embarazo clínico después de 12 meses o más de relaciones sexuales no protegidas.[1] Las técnicas de alta complejidad son aquellos procedimientos que incluyen la manipulación de gametos, o embriones humanos para generar el embarazo. Entre las técnicas incluidas se encuentra la fertilización in-vitro (FIV) y la inyección intra-citoplasmática de espermatozoides (ICSI), transferencia de embriones y criopreservación de los mismos, la donación de ovocitos y embriones y el tratamiento de útero subrogado. [1,2]. En el año 2015 se reglamentó el acceso a las técnicas de reproducción humana asistida de alta complejidad en el Uruguay. La Ley Nº 19.167/013, los Decretos Nº 84/015 y N° 46/017, reglamentaron la financiación de las mencionadas técnicas.[3] Para el acceso a la cobertura las pacientes deberán tener una edad mayor de 18 y menor de 40, y deben cumplir con los criterios establecidos en la normativa de cobertura financiera.[2] El 17 de Agosto del 2020, mediante Decreto del Poder Ejecutivo se modifica la accesibilidad a las técnicas de Reproducción Humana Asistida, eliminado el incremento de los copagos según número de intentos, permitiendo un mayor alcance de la población objetivo.[4] Son tres las clínicas de fertilidad habilitadas en 2015 por el Ministerio de Salud Pública, a realizar procedimientos financiados por el Fondo Nacional de Recursos; Clínica Suizo Americana (CSA), Centro de Esterilidad Montevideo (CEM), y Centro de Reproducción Humana ( CERHIN). OBJETIVOS: Conocer el resultado de las técnicas de reproducción humana asistida de alta complejidad financiadas por el Fondo Nacional de Recursos desde el inicio de la cobertura. METODOLOGÍA: El presente es un estudio observacional retrospectivo, de una cohorte de pacientes usuarias de los procedimientos de RHA, basados en registros secundarios del Fondo Nacional de Recursos y Ministerio de Salud. De acuerdo a los objetivos específicos planteados se consideraron dos puntos de corte diferentes: para el 1er objetivo se tomaron aquellas mujeres incluidas en la cobertura financiera de las técnicas de reproducción humana asistida, desde el inicio de la cobertura en abril de 2015 hasta el 31 de diciembre de 2019. Para el análisis del segundo objetivo se evaluaron los nacidos vivos producto de las técnicas de reproducción humana asistida realizadas hasta el 31 de marzo de 2019 (Nacidos Vivos hasta diciembre de 2019). RESULTADOS: Caracterizar a las pacientes que hicieron uso del programa de RHA desde el inicio de cobertura hasta diciembre de 2019. La caracterización será en función de la edad, clínica de fertilidad, departamento de procedencia y tipo de prestador. CONCLUSIONES: Hasta diciembre de 2019 un total de 3.264 mujeres hicieron uso de las prestaciones financiadas por el Fondo Nacional de Recursos de técnicas de Reproducción Humana Asistida de alta complejidad, atribuyéndose a marzo de 2019 como resultado un total de 1.008 Nacidos Vivos producto de las técnicas financiadas (nacidos vivos hasta diciembre 2019). El corrimiento de la edad de inicio de la maternidad, ha hecho a las técnicas de Reproducción Asistida una opción en el tratamiento de la infertilidad de las parejas. Año a año, se incluyen aproximadamente 700 mujeres a la cobertura por el FNR, la actual disminución de los copagos y la extensión de edad de maternidad proyectan un número de mujeres a financiar creciente. La edad media de presentación ha variado en función del marco normativo, presentado al 2019 un promedio de 36,19 años (el más bajo desde el inicio de la cobertura), la modificación en la edad de presentación seguramente influya en resultados futuros tanto para gestaciones como para Nacidos Vivos. La evolución temporal muestra anualmente una tendencia creciente en la elección de una de las clínicas, observándose la siguiente distribución entre las clínicas con los siguientes porcentajes de distribución: CEM (año 2015 el 64.4%, en 2019 80,4%), CSA (año 2015 22,7%, en 2019 14.5%), Cerhin (año 2015 12.9%, en 2019 5.1%). La mayoría de las transferencias embrionarias evaluadas pertenecieron a transferencias en fresco (60.7%), superando el número de transferencias con embriones criopreservados. Los resultados mostraron una mayor tasa de parto en transferencias de criopreservados respecto a transferencias en fresco, en todos los grupos etarios e independiente de las clínicas de RHA esto debido a las mejores condiciones del endometrio al momento de la implantación. La tasa de parto en relación a la transferencia cae con el aumento de la edad materna, siendo el grupo menor a 35 años el que presentó mejores resultados. En los grupos etarios analizados Uruguay se presenta con una Tasa de parto por transferencia global mejor a la reportada por el resto de los países (según el registro de Red Lara). Respecto a las clínicas de Reproducción Asistida en las categorías analizadas, una de las clínicas presentó mejores tasas de parto por transferencias independiente de la edad materna con valores puntuales favorables en todos los rangos etarios analizados. Los resultados de Ovodonación obtenidos son similares a los obtenidos para ovocitos propios, siendo imputados el 22% de las transferencias realizadas y el 21% del total de los nacidos vivos adjudicables a las técnicas de reproducción Asistida. En un total de 1.008 nacimientos, se presentó una proporción de embarazos múltiples de 20,1% con variabilidad entre clínicas (19,3%-26,2%).

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Abstract This study aimed to evaluate different concentrations of growth and differentiation factor-9 (GDF-9) on the development and maintenance of equine preantral follicle morphology during short-term in vitro culture. Ovaries (n=5) from five mares were collected from a local slaughterhouse and transported to the laboratory, where nine fragments (5x5x1mm) were procured from each ovary. One fragment from each was immediately fixed and submitted for histological analysis (control group; D0). The other eight fragments were cultured in situ for two (D2) or six (D6) days in MEM+ or MEM+ supplemented with GDF-9 at different concentrations (i.e., 50, 100 and 200 ng/mL the GDF-9). After culturing with different concentrations of GDF-9 for 2 or 6 days, the fragments were processed for histological analysis. After two days of cultivation, we observed an increase in the percentage of developing follicles for 0 (MEM+), 50, 100 and 200 ng/mL GDF-9 compared to control (D0; P<0.05). When we evaluated all treatments that preserved follicular integrity, the GDF-9 concentration of 100 ng/mL presented results superior to those of the other cultures (P<0.05). While, at six days of culture, the concentration of 200 ng/mL of GDF-9 appeared to be more efficient in providing development compared to MEM+ (P<0.05). The percentage of morphologically intact follicles in the 6 days culture samples treated with 50 ng/mL of GDF-9 indicated that this concentration was effective in maintaining the integrity of the follicle (P<0.05). We conclude, therefore, that graduated GDF-9 addition to the medium ensure follicular development and is sufficient maintain the architecture.

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This study evaluated a powdered coconut water solution (ACP 406®) as a base culture medium on the in vitro survival and development of in situ goat preantral follicles. The ovarian fragments were either immediately fixed in Carnoy solution (non-cultured control) or individually cultured for 2 or 6 days. The following culture media (all containing 100 μg/mL penicillin and 100 μg/mL streptomycin) were evaluated: α-MEM (α-MEM alone, without additional supplementation); α-MEM+ (supplemented α-MEM); ACP (ACP®406 alone); or ACP+ (supplemented ACP®406). Additional supplementation includes: 1.25 mg/mL bovine serum albumin, 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, 2 mM glutamine, and 2 mM hypoxanthine. The endpoints (i) follicular morphology; (ii) development; (iii) estradiol production; and (iv) reactive oxygen species (ROS) were recorded. Data were analyzed using chi-square, Turkey, t-test or One-Way ANOVA. Differences were considered significant when P 0.05) among treatments. Overall, all treatments had lower primordial follicles (P 0.05) among the treatments. Likewise, no differences (P > 0.05) were observed for ROS production and follicular and oocyte diameters among treatments. Therefore, ACP+ has the equivalent efficiency to MEM+ in maintaining the survival and development of goat preantral follicles, representing an alternative plant-based low-cost culture medium for in vitro culture.

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This study evaluated a powdered coconut water solution (ACP 406®) as a base culture medium on the in vitro survival and development of in situ goat preantral follicles. The ovarian fragments were either immediately fixed in Carnoy solution (non-cultured control) or individually cultured for 2 or 6 days. The following culture media (all containing 100 μg/mL penicillin and 100 μg/mL streptomycin) were evaluated: α-MEM (α-MEM alone, without additional supplementation); α-MEM+ (supplemented α-MEM); ACP (ACP®406 alone); or ACP+ (supplemented ACP®406). Additional supplementation includes: 1.25 mg/mL bovine serum albumin, 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, 2 mM glutamine, and 2 mM hypoxanthine. The endpoints (i) follicular morphology; (ii) development; (iii) estradiol production; and (iv) reactive oxygen species (ROS) were recorded. Data were analyzed using chi-square, Turkey, t-test or One-Way ANOVA. Differences were considered significant when P < 0.05. At day 2 of culture, a greater (P < 0.05) percentage of morphologically normal follicles was observed between ACP+ and ACP treatments. Moreover, at day 2 of culture, no hormonal difference (P < 0.05) was observed between ACP+ and both α-MEM treatments. At day 6 of culture when ACP and α-MEM treatments were compared the percentage of healthy follicles were similar (P > 0.05) among treatments. Overall, all treatments had lower primordial follicles (P < 0.05) accompany by greater developing follicles (P < 0.05) percentages than non-cultured control treatment, indicating primordial follicle activation. However, at day 6 of culture, the percentage of primordial follicle development were similar (P > 0.05) among the treatments. Likewise, no differences (P > 0.05) were observed for ROS production and follicular and oocyte diameters among treatments. Therefore, ACP+ has the equivalent efficiency to MEM+ in maintaining the survival and development of goat preantral follicles, representing an alternative plant-based low-cost culture medium for in vitro culture.(AU)

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SummaryThis study aimed to compare the efficiency of different incubation systems for in vitro embryo production in bovine. Oocytes/embryos were cultured in three incubators: conventional - CONV, mini bench - MINI and portable - PORT. After in vitro maturation (IVM), oocytes were verified for maturation rate. The remaining structures were submitted to in vitro fertilization and culture to verify cleavage (day 2) and blastocyst (day 7) rates. Reactive oxygen species (ROS) were evaluated in post-IVM oocytes and embryos (days 2 and 7) using arbitrary fluorescence units (AFUs). No significant difference (P>0.05) was observed for maturation rate. The CONV system (74.0%) produced the highest cleavage rate (P0.05) to MINI (65.0%). The same pattern and differences were observed for blastocyst rate: CONV (33.3%), MINI (32.3%) and PORT (21.9%). ROS levels were not different (P>0.05) in post-IVM oocytes: CONV (35.6±4.5), MINI (29.4±4.0) and PORT (35.6±4.5). For day-2 embryos, ROS levels were higher (P0.05) was observed in blastocysts. In conclusion, although it produced high ROS levels at day 2 of culture, the MINI system was as efficient as the CONV system for blastocyst production. This option may be an interesting and economical for the in vitro embryo industry.

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OBJECTIVES: Essure® has been tested as an alternative treatment for hydrosalpinx before embryo transfer (ET) in women undergoing assisted reproduction techniques. However, the persistence of a foreign body inside the uterine cavity might have a negative impact on the outcome of pregnancy. The present systematic review aimed at identifying, appraising and summarizing the available evidence regarding the effectiveness and safety of using Essure prior to ET for women with hydrosalpinx. METHODS: We searched for studies in PubMed, Scopus, CENTRAL, Web of Science and ClinicalTrials.gov and the reference lists of eligible studies. All studies including at least 10 women with hydrosalpinx who received Essure, any other intervention or no treatment prior to ET were considered eligible. Study selection, data extraction and evaluation of the risk of bias were performed independently by two authors. Study outcomes were miscarriage per clinical pregnancy, singleton preterm birth per singleton live birth and live birth/ongoing pregnancy and clinical pregnancy per ET. The pooled results for each outcome and intervention were summarized as proportions with their respective 95% CIs, using a random-effects model. RESULTS: Our electronic search of databases was performed on 7 November 2015, and 26 studies with 43 study arms were considered eligible: eight study arms evaluating Essure; seven assessing tubal aspiration; seven appraising effects of no treatment; 12 evaluating salpingectomy; two assessing tubal division; and seven evaluating tubal occlusion. When compared with women who had no intervention, women with Essure had a higher clinical pregnancy rate per ET (36% (95% CI, 0-43%) vs 13% (95% CI, 9-17%)). When compared with women who had other interventions, women with Essure had a higher miscarriage rate per clinical pregnancy (38% (95% CI, 27-49%) vs 15% (95% CI, 10-19%)). CONCLUSIONS: The available evidence suggests that, although Essure prior to ET in women with hydrosalpinx improves the chance of achieving a clinical pregnancy compared with no intervention, it is associated with a higher rate of miscarriage when compared with the other interventions. Although this evidence is based on observational studies, we believe that salpingectomy should be the first option for women who are eligible for videolaparoscopy. However, it is still premature to make recommendations for women who are not eligible for surgery, and randomized controlled trials are needed to clarify which is the best treatment alternative in such a scenario. Copyright © 2016 ISUOG. Published by John Wiley & Sons Ltd.

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A partir de los registros de transferencia de embriones producidos in vitro de tantos años de la central genética en el municipio de San Pedro de los Milagros Antioquia – Colombia, se buscó retrospectivamente si existía unarelación sobre la cantidad de ovocitos y embriones viables producidos por las donadoras. Dependiendo de la raza,se buscó determinar si existe una influencia de la donante. Las variables analizadas fueron: el toro, la raza, el lote, la historia reproductiva de las donadoras, las cuales se encontraban en iguales condiciones de manejo, tanto sanitarias y nutricionales. La evaluación de los resultados se realizó mediante el modelo estadístico de Chi-cuadrado.Para la comparación entre el porcentaje de ovocitos viables y el porcentaje de embriones producidos se realizó unaprueba de análisis de varianza (ANOVA) y una prueba de Fisher con un intervalo de confianza del 95% y un valorde p>0.05. Se encontraron diferencias estadísticamente significativas en la producción de embriones (40% vs 29%), taurus e indicus, respectivamente. Se concluye, que las donadoras taurus producen mayor número de embrionesque las indicus.

From the many years of transfer records of embryos produced in vitro at the genetics center in the municipality ofSan Pedro de los Milagros, Antioquia, Colombia, we examined retrospectively if there was a relationship betweenthe number of viable oocytes and embryos produced by the donors. Depending on the race, we sought to determine whether there is an influence by the donor. The variables analyzed were: the bull, race, batch, and reproductive history of the donors, all of which were in similar health and nutritional conditions. The evaluation of the results was carried out with the Chi- Square test statistical model. In order to compare between the percentage of viable oocytes and the percentage of embryos, an analysis of variance (ANOVA) was performed as well as a Fischer test, with a confidence interval of 95% and a p-value of p>0.05. Statistically significant differences were found in the production of embryos (40% vs 29%) taurus and indicus, respectively. The conclusion is that taurus donors produce greater numbers of embryos than indicus donors.

A partir dos registros de transferência de embriões produzidos in vitro nos muitos anos da central genética nomunicípio de San Pedro de los Milagros, Antioquia- Colômbia procuramos retrospectivamente se há uma relaçãoentre a quantidade do número de oócitos viáveis e embriões produzidos por doadores. Dependendo da raça,buscou-se determinar se há uma influência do doador. As variáveis analisadas foram: o touro, a raça, o lote, a história reprodutiva dos doadores, os quais encontravam-se nas mesmas condições de manejo, saúde e nutrição. A avaliação dos resultados foi realizada utilizando o modelo de teste estatístico Chi-quadrado. Para a comparação entrea porcentagem de oócitos viáveis e a porcentagem de embriões produzidos foram realizados um teste pela análisede variância (ANOVA) e um teste de Fisher, com intervalo de confiança de 95% e um valor de p>0.05. Foramencontradas diferenças estatisticamente significativas na produção de embriões (40% vs 29%), taurus e indicus,respectivamente. A conclusão é que os doadores taurus produzem um número maior de embriões que os doadoresindicus.

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La producción in vitro de embriones (PIVE) en equinos es una técnica que genera incrementos en la eficienciareproductiva de especímenes de alto valor genético y comercial. A pesar de todos los estudios que se hanrealizado, la PIVE es una técnica con acceso y un éxito limitado, debido principalmente a las dificultades paraalcanzar tasas de fertilización eficientes mediante fertilización in vitro convencional, al igual que por las altasnecesidades en recursos técnicos y económicos para la fertilización mediante inyección intracitoplasmáticade espermatozoides (ICSI), entre otras limitaciones existentes para el desarrollo embrionario in vitro. En elmundo, se han evaluado múltiples alternativas para el mejoramiento de la PIVE en equinos, desde el uso dediferentes medios, fluidos, células, y moléculas, hasta la utilización de métodos alternativos para mejorar lastasas de fertilización como la ICSI en asociación de sustancias como el polivinilalcohol. Esta revisión exploralos avances investigativos y las expectativas futuras de aplicación de la técnica de PIVE en equinos.

The in vitro embryo production (PIVE) in horses is a technique that generates increases in reproductive efficiencyof high genetic and commercial value animals. Despite all the studies that has been made, PIVE is still atechnique with limited access and success, mainly due to difficulties to achieve efficient rates of fertilizationthrough conventional in vitro fertilization, as well as by the high needs in technical and economic resources for fertilization using intracytoplasmic sperm injection (ICSI), among other limitations on in vitro embryonic development. In the world there have been evaluated multiple alternatives for improving the PIVE in horses: from the use of different means, fluids, cells and molecules, to the use of alternative methods to improve therates of fertilization through ICSI with the use of substances such as polyvinylalcohol. This review explores the research progress and future expectations for the application of the PIVE technique in horses.

A produção de embriões eqüinos in vitro (PIVE) é uma técnica que aumenta eficiência reprodutiva em espéciesde alto valor genético e comercial. Apesar de todos os estudos realizados, a PIVE é uma técnica de acesso eêxito limitados, principalmente devido à dificuldade de se alcançar taxas de fertilização eficientes diante dafertilização in vitro convencional, grande necessidade de recursos financeiros e técnicos para a fertilizaçãopor injeção intracitoplasmática de espermatozóide (ICSI), entre outras limitações para o desenvolvimentoembrionário in vitro. Em todo mundo foram avaliadas várias alternativas para a melhoria da PIVE em eqüinos,desde a utilização de diferentes meios, fluidos, células e moléculas até a utilização de métodos alternativos paramelhorar as taxas de fertilização como a ICSI em associação com outras substâncias como o álcool polivinílico.Essa revisão aborda o progresso da pesquisa e as perspectivas futuras de aplicação da técnica PIVE em eqüinos.

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Temperature and rainfall were analyzed daily during six years to evaluate their influence on in vitro production of bovine embryos. Weekly replications (n=480) were performed on 14,778 ovaries collected at slaughterhouses. Cumulus oocyte complexes (n=19,180) were fertilized with a pool of Bos taurus taurus semen in one incubator with 5 percent CO2. Presumable zygotes were cultured in gasified plastic bags with 5 percent CO2, 5 percent O2, and 90 percent N2. In the first year, cleavage and embryo yield were 60.3 percent and 15.6 percent, respectively, being lower (P<0.05) than in the following years. Average cleavage rates were always lower in winter (P<0.0001), thus producing less embryos. Winter climatic conditions had a negative influence on in vitro production, when cleavage and embryo yield declined, possibly because of reduced availability and growth of native pasture.(AU)

A temperatura e a precipitação pluviométrica foram analisadas diariamente, durante seis anos, para avaliar sua influência sobre a produção in vitro de embriões bovinos. As repetições semanais (n=480) foram realizadas com 14.778 ovários coletados em matadouros. Os oócitos (n=19.180) foram maturados em estufa com atmosfera com controle de temperatura e umidade saturada com 5 por cento de CO2 e, após 20h, foram fecundados com sêmen de Bos taurus taurus e mantidos sob as mesmas condições de atmosfera da maturação. Os zigotos foram cultivados em placas de quatro poços em bolsas gaseificadas com 5 por cento de CO2, 5 por cento de O2 e 90 por cento de N2, à temperatura de 39ºC e umidade saturada. No primeiro ano, a taxa clivagem (60,3 por cento) e a produção de embriões (15,6 por cento) foram inferiores (P<0,05) aos demais anos. As taxas de clivagem foram sempre menores no inverno (P<0,0001). As condições climáticas no inverno tiveram influência negativa sobre a produção in vitro de embriões bovinos e houve diminuição nos índices de clivagem e produção de blastocistos, possivelmente devido à reduzida disponibilidade e crescimento da pastagem nativa.(AU)

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Temperature and rainfall were analyzed daily during six years to evaluate their influence on in vitro production of bovine embryos. Weekly replications (n=480) were performed on 14,778 ovaries collected at slaughterhouses. Cumulus oocyte complexes (n=19,180) were fertilized with a pool of Bos taurus taurus semen in one incubator with 5 percent CO2. Presumable zygotes were cultured in gasified plastic bags with 5 percent CO2, 5 percent O2, and 90 percent N2. In the first year, cleavage and embryo yield were 60.3 percent and 15.6 percent, respectively, being lower (P<0.05) than in the following years. Average cleavage rates were always lower in winter (P<0.0001), thus producing less embryos. Winter climatic conditions had a negative influence on in vitro production, when cleavage and embryo yield declined, possibly because of reduced availability and growth of native pasture.

A temperatura e a precipitação pluviométrica foram analisadas diariamente, durante seis anos, para avaliar sua influência sobre a produção in vitro de embriões bovinos. As repetições semanais (n=480) foram realizadas com 14.778 ovários coletados em matadouros. Os oócitos (n=19.180) foram maturados em estufa com atmosfera com controle de temperatura e umidade saturada com 5 por cento de CO2 e, após 20h, foram fecundados com sêmen de Bos taurus taurus e mantidos sob as mesmas condições de atmosfera da maturação. Os zigotos foram cultivados em placas de quatro poços em bolsas gaseificadas com 5 por cento de CO2, 5 por cento de O2 e 90 por cento de N2, à temperatura de 39ºC e umidade saturada. No primeiro ano, a taxa clivagem (60,3 por cento) e a produção de embriões (15,6 por cento) foram inferiores (P<0,05) aos demais anos. As taxas de clivagem foram sempre menores no inverno (P<0,0001). As condições climáticas no inverno tiveram influência negativa sobre a produção in vitro de embriões bovinos e houve diminuição nos índices de clivagem e produção de blastocistos, possivelmente devido à reduzida disponibilidade e crescimento da pastagem nativa.

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