Desarrollo de un sistema económico para la vitrificación de embriones de conejo

Abstract

[ES] Uno de los soportes más utilizados actualmente en la vitrificación de óvulos y embriones es el denominado Cryotop. Este soporte permite altos ratios de enfriamiento debido al pequeño volumen de la solución de vitrificación empleada, si bien, su precio ronda los 10 euros. El objetivo de este trabajo es comparar el Cryotop con un asa de siembra de plástico para la vitrificación de embriones de conejo. Tras la inseminación, a las 72 horas se recuperarán embriones en estadios de mórula o blastocisto temprano que serán sometidos a vitrificación, en una solución de 20% de Etilenglicol y un 20% de Dimetlsulfóxido, en los 2 soportes anteriormente descritos: Cryotop y asa de siembra. Por una parte los embriones serán desvitrificados y cultivados in vitro en una solución TCM199 con un 10% de suero a 38.5ºC y humedad saturada durante 48 horas. Se determinará la capacidad de desarrollo a blastocisto escapando o escapado. En un segundo experimento, embriones de ambos grupos experimentales serán transferidos mediante laparoscopia para evaluar la tasa de implantación a los catorce días tras la inseminación y el número de nacidos vivos a parto.

[EN] Over the last several decades there have been many studies in order to develop efficient carriers that produce minimum damage over the cells. Cryotop is one of the more currently employed carriers for oocyte and embryo vitrification due to the high cooling rates achieved because it reduces the volume of vitrification, however, its price is around 21€ per each. The aim of this work is to compare between Cryotop and plastic inoculating loop for rabbit embryo vitrification. Briefly, 72 hours after insemination embryos were recovered in morulae or early blastocyst stage and were vitrified using vitrification solution consisting of 20% (vol/vol) ethylene glycol and 20% (vol/vol) dimethyl sulfoxide. Then embryos were loaded in Cryotop or inoculating loop. On the one hand, embryos were devitrified and cultured in vitro during 48 hours in medium TCM199 containing 10% serum at 38,5ºC and humidified atmosphere. It was evaluated the development of embryos until hatching. On the other hand, embryos from both experimental groups were transferred using the laparoscopic technique to evaluate implantation rate at fourteen days after insemination and offspring rate at birth. There were no differences between Inoculating loop and Cryotop under in vivo and in vitro culture conditions. Therefore, inoculating loop is a suitable method for replacing Cryotop.