Oocyte Selection for in Vitro Embryo Production in Livestock Species: Non-Invasive Approaches for the New Challenges of Oocyte Competence

Águila, Luis; Treulén, Favián; Therrien, Jacinthe; Felmer, Ricardo; Valdivia, Martha; C, Smith Lawrence

doi:10.20944/preprints202010.0453.v1

Cited by 2 publications

(1 citation statement)

References 116 publications

(145 reference statements)

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“…El contenido se volcó a una placa Petri de 90 x 15 mm con una cuadrícula para que la búsqueda de los CCOs resultara más sencilla y se seleccionaron los CCOs inmaduros bajo el estereomicroscopio en la cabina de flujo laminar. Se seleccionaron sólo aquellos CCOs que tenían tres, cuatro o más capas de células del cumulus compactas alrededor del ovocito y cuyo citoplasma fuera lo más homogéneo posible (Aguila et al, 2020;Telfer et al, 2020). Los CCOs seleccionados se pasaron a una placa Petri de 35 x 10 mm con 2 ml de OCM suplementado con una pipeta Pasteur de vidrio estirada a la llama acoplada a una cánula de silicona.…”

Section: Recogida De Ovarios Selección De Ovocitos Y Maduración In Vi...unclassified

Efecto De Las Secreciones De Organoides De Endometrio Humano Sobre El Cultivo in Vitro De Embriones Bovinos

Masegosa Domínguez,

Romero-Aguirregomezcorta,

Juan

et al. 2024

An. Vet. Murcia

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Over the last 50 years, the fertility of high-producing dairy cows has declined due to the intensity of genetic selection. To overcome this obstacle, assisted reproductive techniques have been used. However, in vitro embryo production is a relatively inefficient process, with maximum blastocyst yield rates of around 35%. To overcome this problem, it was hypothesised that by adding endometrial organoid secretions to the culture medium, embryos produced in vitro would be exposed to a more physiological environment and the yield of the technique would increase. For this purpose, bovine oocytes were collected from a local abattoir and matured in vitro, fertilised in vitro and cultured in vitro. The embryo culture medium was supplemented at 1% (v/v) with i) human endometrial organoid secretions (as bovine secretions are not available), ii) Boretto culture medium, and ii) matrigel drops with Boretto culture medium, as procedural controls. In addition, a control group without any supplementation was included. Embryo development was assessed at days 2, 7 and 8 of culture. The percentage of blastocysts at day 7 and day 8 post insemination was significantly higher (P<0.05) when Boretto culture medium was added. In addition, this treatment increased embryo development kinetics at day 7 post insemination, and blastocyst quality as assessed by total cell number, with 85.8% more than the control. Although human endometrial secretions have not given the expected result, it would be necessary to test whether bovine endometrial-specific organoid secretions, at different concentrations, could increase the yield of bovine embryo production in vitro. Durante los últimos 50 años, la fertilidad de las vacas lecheras de alta producción ha disminuido debido a la intensidad de la selección genética. Para superar este obstáculo, se ha recurrido a las técnicas de reproducción asistida. Sin embargo, la producción de embriones in vitro es un proceso relativamente ineficiente, con tasas máximas de blastocistos obtenidos de alrededor del 35%. Para solventar este problema, se planteó la hipótesis de añadir al medio de cultivo secreciones de organoides de endometrio, así, los embriones producidos in vitro estarían expuestos a un ambiente más fisiológico y el rendimiento de la técnica aumentaría. Para ello, se llevó a cabo la recolección de ovocitos bovinos procedentes de un matadero local, y se procedió a su maduración in vitro, fecundación in vitro y cultivo embrionario in vitro. El medio de cultivo embrionario se suplementó al 1% (v/v) con i) secreciones de organoides de endometrio humano (ya que no existe disponibilidad de secreciones bovinas), ii) medio de cultivo Boretto, y ii) gotas de matrigel con medio de cultivo Boretto, como controles de procedimiento. Además, se incluyó un grupo control sin ningún suplemento. Se evaluó el desarrollo embrionario a los días 2, 7 y 8 de cultivo. El porcentaje de blastocistos a día 7 y a día 8 post inseminación fue significativamente mayor (P<0.05) cuando se añadió el medio de cultivo Boretto. Además, este tratamiento incrementó la cinética de desarrollo de los embriones a día 7 post inseminación, y la calidad de los blastocistos evaluada por el número total de células, con un 85.8% más que el control. Aunque las secreciones de endometrio humano no han dado el resultado esperado, sería necesario comprobar si las secreciones de organoides específicas de endometrio bovino, a distintas concentraciones, podrían incrementar el rendimiento de la producción in vitro de embriones bovinos.

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Section: Recogida De Ovarios Selección De Ovocitos Y Maduración In Vi...unclassified

Efecto De Las Secreciones De Organoides De Endometrio Humano Sobre El Cultivo in Vitro De Embriones Bovinos

Masegosa Domínguez,

Romero-Aguirregomezcorta,

Juan

et al. 2024

An. Vet. Murcia

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Simple and Efficient Chemically Defined In Vitro Maturation and Embryo Culture System for Bovine Embryos

Arias

Vargas

Gallardo

et al. 2022

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Supplementation of the culture media for in vitro production (IVP) of bovine embryos with fetal bovine serum (FBS) is associated with inconsistent outcomes. The present study sought to replace FBS and BSA by insulin-like growth factor 1 (IGF1), fibroblast growth factor 2 (FGF2) and epidermal growth factor (EGF). In Experiment 1, absence of FBS from maturation medium (MM) did not affect the rate of in vitro maturation, as assessed by the extrusion of the first polar body. However, when gonadotropins and FBS were removed from the MM, the maturation rate was significantly reduced even in the presence of growth factors. Therefore, gonadotropin-supplemented MM medium was established as the base medium for the defined maturation condition. In Experiment 2, the addition of growth factors to gonadotropin-supplemented MM medium supported similar maturation (~90%) compared to the undefined condition (FBS-carrying). In Experiment 3, the addition of growth factors to embryo culture medium showed similar in vitro competence compared to the undefined (FBS) control. In Experiment 4, completely defined conditions (absence of FBS and BSA during in vitro maturation and embryo culture) were tested. A higher cleavage was observed with FGF2 (86%) compared to EGF (77%) and the FBS control (77%), but similar blastocyst rates were observed for FGF2 (24%), EGF (19%) and the FBS control (25%). Embryo quality was similar among groups. Finally, post-thawing survival was higher for FGF2 (94%) compared to the FBS control (77%). Thus, we report a simple defined IVP system for bovine species that generates developmental outcomes and embryos of similar quality than those produced under conditions containing FBS.

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Oocyte Selection for in Vitro Embryo Production in Livestock Species: Non-Invasive Approaches for the New Challenges of Oocyte Competence

Cited by 2 publications

References 116 publications

Efecto De Las Secreciones De Organoides De Endometrio Humano Sobre El Cultivo in Vitro De Embriones Bovinos

Efecto De Las Secreciones De Organoides De Endometrio Humano Sobre El Cultivo in Vitro De Embriones Bovinos

Simple and Efficient Chemically Defined In Vitro Maturation and Embryo Culture System for Bovine Embryos

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