Improvements to In Vitro Culture Media for Use in Bovine IVF

Vh, Do; Walton, Simon; Aw, Taylor-Robinson

doi:10.15744/2348-9790.4.205

Cited by 4 publications

(3 citation statements)

References 51 publications

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“…La producción in vitro de embriones (PIV) y las tecnologías asociadas comenzaron a desarrollarse en el ganado bovino en la década de los 80 y han mostrado importantes avances en los últimos años (Ferré et al, 2020). La producción in vitro implica cuatro pasos principales en secuencia: obtención de ovocitos y maduración in vitro (MIV); selección espermática; fecundación in vitro (FIV); y cultivo in vitro (CIV) de embriones tempranos preimplantatorios (Vh et al, 2016). Todos estos procesos pretenden "imitar" in vitro lo que ocurre en condiciones in vivo en el tracto reproductor de la hembra.…”

Section: Introductionunclassified

“…En la reproducción asistida del ganado bovino, el diseño y la preparación de medios de cultivo in vitro han sido fundamentales para conseguir el desarrollo adecuado de ovocitos y embriones (Vh et al, 2016). En términos de eficiencia, aproximadamente el 90% de los ovocitos inmaduros experimentan una maduración nuclear exitosa y avanzan de la profase I a la metafase II durante la maduración in vitro .…”

Section: Introductionunclassified

“…Los medios de CIV carecen de componentes esenciales que permiten el correcto funcionamiento de las vías fisiológicas necesarias para el desarrollo de embriones bovinos (Vh et al, 2016). Para solventar este problema, comúnmente se ha utilizado el suero fetal bovino (SFB) en el medio de MIV y de CIV, para aumentar la eficiencia de la maduración de los ovocitos y el cultivo embrionario.…”

Section: Introductionunclassified

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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: Introductionunclassified

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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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Cryopreservation of in vitro-produced bovine embryos by vitrification: In pursuit of a simplified, standardized procedure that improves pregnancy rates to promote cattle industry use

Huong¹,

Taylor-Robinson

2020

BIOTEC ANIM HUSB

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The goal of cryopreservation is to retain the original stage of gametes and embryos after they have endured cooling and warming. Slow freezing is a standard method for in vivo-derived bovine embryo cryopreservation, threefifths of such embryos being frozen by this method globally. However, it is evident that slow freezing is not efficient for cryopreserving in vitro-produced bovine embryos. Hence, only one-third of in vitro-produced bovine embryos are cryopreserved. Vitrification is a preferred method for storage of human embryos; consequently, it has been explored as a novel means to store in vitro-produced bovine embryos, for which it shows considerable promise as an alternative to slow freezing. This is due to several reasons: vitrification is often less time-consuming than slow freezing; it does not need expensive slow rate freezing machines; and it has been proven to have comparatively higher survival rates. Yet, in the cattle industry vitrification continues to present shortcomings, such as possible toxicity of vitrification solutions and failure to standardize methods, which pose a challenge for its application to in vitro-produced bovine embryos. Therefore, determining the most suitable procedure is crucial to make vitrification more practical in commercial settings.

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Improvement of bovine in vitro embryo production by fetal calf serum and cysteamine supplementation and investigation of freezability

Sandal

Şenlikçi

Elgün

et al. 2020

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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The aim of this study was to investigate the effects of cysteamine (Cys) and fetal calf serum (FCS) in synthetic oviduct fluid (SOF) and charles and rosenkrans (CR1aa) on the bovine in vitro embryo production and its ability of freeze. The oocytes were divided into two groups and allowed to mature in TCM-199, with and without cysteamine. They were divided into 4 subgroups according to whether they contain Cys and FCS in 2 different culture media (SOF and CR1aa). Accordingly, 8 groups were formed as SOF+FCS+Cys, SOF+Cys, SOF+FCS, SOF, CR1aa+FCS+Cys, CR1aa+Cys, CR1aa+FCS and CR1aa. Embryos were cultured for 7-9 days and were frozen using vitrification method. Development of embryos was observed during the first 24 hr post-thaw period. The highest rate of compact morula was 20% in SOF+FCS+Cys and 19.4% in CR1aa+FCS+Cys. The rate of blastocyst in SOF+FCS and CR1aa+FCS were found 17.9% and 15.4% respectively and the difference between groups was statistically significant (P<0.05). During the 12 hr vitality assessment, the highest rate was determined in SOF+FCS (47%). The results indicate that FCS has a positive effect in reaching compact morula and blastocyst regardless of the presence of the culture medium or antioxidant used. There was no statistically difference between the values of post-thaw embryos.

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Improvements to In Vitro Culture Media for Use in Bovine IVF

Cited by 4 publications

References 51 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

Cryopreservation of in vitro-produced bovine embryos by vitrification: In pursuit of a simplified, standardized procedure that improves pregnancy rates to promote cattle industry use

Improvement of bovine in vitro embryo production by fetal calf serum and cysteamine supplementation and investigation of freezability

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