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Co-culture with granulosa cells improve the in vitro maturation ability of porcine immature oocytes vitrified with cryolock.

Authors :
Casillas F
Teteltitla-Silvestre M
Ducolomb Y
Lemus AE
Salazar Z
Casas E
Betancourt M
Source :
Cryobiology [Cryobiology] 2014 Oct; Vol. 69 (2), pp. 299-304. Date of Electronic Publication: 2014 Aug 23.
Publication Year :
2014

Abstract

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.<br /> (Copyright © 2014 Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
1090-2392
Volume :
69
Issue :
2
Database :
MEDLINE
Journal :
Cryobiology
Publication Type :
Academic Journal
Accession number :
25152224
Full Text :
https://doi.org/10.1016/j.cryobiol.2014.08.004