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Viability, maturation and embryo development in vitro of immature porcine and ovine oocytes vitrified in different devices
Authors:Fernández-Reyez Filiberto  Ducolomb Yvonne  Romo Salvador  Casas Eduardo  Salazar Zayil  Betancourt Miguel
Affiliation:Departamento de Producción Agrícola y Animal, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Xochimilco, Calzada del Hueso 1100, 04960 DF, Mexico. freyes@correo.xoc.uam.mx
Abstract:This study was designed to evaluate the effects of vitrification on immature porcine and ovine oocytes, collected at a slaughterhouse, by performing vitrification in devices with different volumes. Viability was evaluated both before and after vitrification and maturation. Immediately after warming, the percentage of viable pig oocytes was 81% regardless the type of device, while in the control (after oocyte selection) was 95%. The viability of matured pig oocytes after warming, vitrified in beveled edge open straws (BES) was 6%, in small-open-pulled-straw (SOPS) was 17% and in cryotop was 4%, while the viability of the control group was 86%. The viability and maturation results were similar with all devices. Embryo development (ED) was observed in fresh porcine oocytes with 15% 2-8 cell embryos, 7% morulae and 3% blastocysts, and non-embryo cleavage was observed in warmed oocytes. The viability of sheep oocytes immediately after warming averaged 90% in all devices, while that of the control (after oocyte selection) averaged 95%. The viability of warmed oocytes after maturation was: BES 21%, SOPS 30%, cryotop 21% and control group 86%; while maturation values were 11, 21, 34 and 70%, respectively. After vitrification, the highest ED was achieved with ovine oocytes vitrified in SOPS, with 17% morulae development and it was the only device in which blastocysts developed. A direct relationship was observed between viability and actin filament integrity in both species.
Keywords:BES, beveled edge open straw   BSA, bovine serum albumin   COCs, cumulus cell-oocyte-complexes   DPBS, Dulbecco’s phosphate buffered solution   ED, embryo development   EG, ethylene glycol   FCS, fetal calf serum   GV, germinal vesicle   IVF, in vitro fertilization   IVM, in vitro maturation   MII, metaphase II   mTBM, modified Tris-buffered-medium   MTT, thiazolyl blue   NCSU-23, North Caroline State University-23   OPS, open pulled straw   PVA, polyvinyl alcohol   SOPS, small open pulled straw   TCM-199, tissue culture medium 199   TL, tyrode lactate
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