Vitrificacion de blastocitos bovinos producidos in vitro con el método Open Pulled Straw (OPS): Primer reporte
Vitrificacion de blastocitos bovinos producidos in vitro con el método Open Pulled Straw (OPS): Primer reporte
Authors
Silva, M.E.
Berland Olea, Marco
Berland Olea, Marco
Authors
Date
2012-02-27
Datos de publicación:
10.4067/S0301-732X2004000100009
Keywords
Embriones bovinos - Bovino - Vitrificación
Collections
Abstract
El objetivo de este trabajo fue presentar los primeros resultados exitosos de vitrificación de blastocistos bovinos producidos in vitro utilizando la técnica del Open Pulled Straw (OPS) en nuestro país. Para la confección de las OPS, pajuelas plásticas de 0.25 ml fueron estiradas utilizando calor. Los 93 blastocistos seleccionados para la vitrificación fueron depositados individual y secuencialmente en las soluciones de vitrificación 1 y 2 consistentes en TCM199 (20% SFB) adicionado con Etilénglicol y Dimetilsulfoxido (DMSO), por períodos de un minuto y 20 segundos respectivamente. Durante este último período el embrión fue cargado con una micropipeta, en un volumen de 2 µl, depositándose sobre una placa petri desde la cual fue cargado por capilaridad en la pajuela estirada, para luego depositarla directamente en el nitrógeno líquido. El 89% de los blastocistos congelados fueron recuperados post descongelación, de los cuales 54% se encontraban re-expandidos o eclosionados a las 24 horas de cultivo. Al evaluar los embriones a las 72 horas post descongelación se observó un 29% de embriones eclosionados. Aunque levemente inferiores a resultados reportados en la literatura, los nuestros indican la factibilidad de vitrificar exitosamente blastocistos bovinos producidos in vitro utilizando el método de Open Pulled Straw.
The purpose of this work is to present preliminary produced in vitro using the Open Pulled Straw (OPS) method. Open Pulled Straws were made by pulling 0.25 ml inner and outer diameter were reduced to half of their original and deposited individually in vitrification solutions 1 and dimethyl - sulfoxide (DMSO), for 1 minute and 20 µl containing the embryo pulled into the straw by capillarity into liquid Nitrogen. Eighty three blastocysts were recovered after thawing. Fifthy four percent of these were re- expanded or hatched after 24 hours of in vitro culture. After 72 hours of in vitro culture 29% of the blastocysts matured. Even though our results are slightly lower to those reported in the literature, they show that it is possible to successfully vitrify bovine blastocysts produced in vitro using the OPS method.
The purpose of this work is to present preliminary produced in vitro using the Open Pulled Straw (OPS) method. Open Pulled Straws were made by pulling 0.25 ml inner and outer diameter were reduced to half of their original and deposited individually in vitrification solutions 1 and dimethyl - sulfoxide (DMSO), for 1 minute and 20 µl containing the embryo pulled into the straw by capillarity into liquid Nitrogen. Eighty three blastocysts were recovered after thawing. Fifthy four percent of these were re- expanded or hatched after 24 hours of in vitro culture. After 72 hours of in vitro culture 29% of the blastocysts matured. Even though our results are slightly lower to those reported in the literature, they show that it is possible to successfully vitrify bovine blastocysts produced in vitro using the OPS method.