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Cryobiology ; 69(2): 299-304, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25152224

RESUMEN

This study was designed to evaluate the efficiency of two oocyte vitrification-warming procedures using two different devices: Superfine Open Pulled Straws (SOPS) and Cryolock, as well as the effect of the co-culture of vitrified immature oocytes with fresh granulosa cells to improve in vitro maturation (IVM). Immature oocytes were vitrified with two procedures: A) Oocytes were exposed to an increasing concentration of ethylene glycol (EG) from 4% to 35% with 0.5 M trehalose. They then, were loaded in SOPS or Cryolock. For warming, oocytes were exposed to decreasing concentrations of trehalose 0.3, 0.2 and 0.1 M for IVM. B) Oocytes were exposed to two mixtures of EG and dimethylsulfoxide (Me2SO), at 7.5% and 16%, both with 0.4 M of sucrose and then loaded in SOPS or Cryolock and stored in liquid nitrogen. For warming, oocytes were exposed to a single concentration of sucrose 0.5M. After warming, viability was determined; and after 44 h of IVM both viability and meiotic stages were evaluated. The results indicate no significant differences between procedures A and B with SOPS in all maturation stages, reaching a maximum maturation rate of 21%. As to Cryolock, significant differences were observed between both procedures, being procedure B, more efficient with a yield of 38% in MII stage and increased to 49% due to the co-culture with fresh granulosa cells. In conclusion, viability and maturation rates were improved with Cryolock and procedure B with the co-culture system in vitrified immature oocytes.


Asunto(s)
Técnicas de Cocultivo/veterinaria , Criopreservación/veterinaria , Células de la Granulosa/citología , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Oocitos/citología , Vitrificación , Animales , Supervivencia Celular , Células Cultivadas , Técnicas de Cocultivo/métodos , Criopreservación/métodos , Crioprotectores/metabolismo , Dimetilsulfóxido/metabolismo , Glicol de Etileno/metabolismo , Femenino , Técnicas de Maduración In Vitro de los Oocitos/métodos , Porcinos , Trehalosa/metabolismo
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