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Influence of Vitrification on the Chromosomes and Spindles of Porcine Oocytes Matured in Vitro
Zhi-Feng Xiang*, Jin-Zhou Zhang, Pei-Qing Li, De-Fu Zhang1, Shi-Qing Zhang1
Department of Animal Science, Henan Sci-tech College, Xinxiang 453003, China; 1Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China
Abstract: To investigate the influence of vitrification freezing on the chromosomes and spindles of porcine oocyte matured in vitro by using nylon cryoloop. Porcine oocytes were verified with ethylene glycol (EG) singly or EG combined with dimethyl sulphoxide (DMSO) as cryoprotectants， and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine. After frozen porcine oocytes thawed， the microtubulin of the spindles and chromosomes were fixation and stained by immunofluorescent method. Normal morphological oocytes in every groups were used to fertilize in vitro. Comparatively， in protocol of EG combined with DMSO and at ultra-rapid cooling rate， the normal configuration of spindle and chromosomes rate of thawed porcine oocytes was remarkably higher than that of the two protocol of single EG used and EG combined with DMSO， As the same in the rate of oocytes activated. The protocol of EG combined with DMSO as cryoprotectants and with extremely high cooling rate can produce better effect on conservation of chromosomes and spindle configuration in vitrification of porcine oocytes， while the development ability of thawed oocytes are severely affected.
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