ACTA VETERINARIA ET ZOOTECHNICA SINICA

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Effects of Three Vitrification Methods on the Mitochondria Distribution and Damage of Porcine MII-stage Oocytes

DAI Jian-jun1,2, WU Cai-feng1,2, ZHANG Ting-yu1,2, ZHANG Shu-shan1,2, XU Li1,2, ZHANG De-fu1,2*   

  1. (1. Institute of Veterinary Science, Shanghai Academy of Agricultural Sciences, Shanghai 201106,
    China; 2. The Research Group of Animal Genetic Engineering, Shanghai Municipal Key
    Laboratory of Agri-genetics and Breeding, Shanghai 201106, China
    )
  • Received:2012-01-08 Online:2012-10-25 Published:2012-10-25

Abstract: The purpose of this study was to detect the mitochondria distribution and ultrastructure changes that resulted from vitrified porcine MII-stage oocytes with three vitrification techniques. Through transmission electron microscopy (TEM), R-123 (Rhodamine-123) staining and in vitro development, mitochondria distribution and ultrastructure were observed. The result showed that: (1) Given the survival rate of FDA (fluorescein diacetate)-DAPI (4′,6′-diamidino-2-phenylindole dihydrochloride) staining and the cleavage rate after parthenogenetic activation, CLV (Cryoloop vitrification) method (72.00%, 7.22%) got the best results. OPS (Open Pulled Straw,60.00%, 4.85%) and straw (42.22%, 0%) methods were followed by; (2) The rate of normal mitochondria distribution in CLV group (52.24%) was higher than that in the two others groups (OPS, 48.65%; straw, 37.68%) , but there was no significant differences among them (P>0.05); (3) Vague mitochondria in thawed oocytes were observed in the ultrastructure of vitrified oocytes from TEM, and some mitochondrial ridges were reduced or even disappeared. The result indicated that mitochondria was damaged greatly in the freezing process and CLA method could improve the freezing speed and minimize such damages.

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