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RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.


 

Article << Previous     |     Next >>   Contents Vol 19(2)

Effects of vitrification procedures on subsequent development and ultrastructure of in vitro-matured swamp buffalo (Bubalus bubalis) oocytes

Duangjai Boonkusol A B, Tassanee Faisaikarm C, Andras Dinnyes D E, Yindee Kitiyanant A C F

A Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
B Department of Biology, Faculty of Science, Srinakharinwirot University, Bangkok 10110, Thailand.
C Institute of Science and Technology for Research and Development, Mahidol University, Salaya, Nakhon Pathom 73170, Thailand.
D Genetic Reprogramming Team, Agricultural Biotechnology Center, 2100 Godollo, Hungary.
E BioTalentum Ltd, 2100 Godollo, Hungary.
F Corresponding author. Email: scykt@mahidol.ac.th
 
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Abstract

The purpose of the present study was to investigate the effects of two vitrification procedures on developmental capacity and ultrastructural changes of matured swamp buffalo oocytes. In vitro-matured oocytes were vitrified by using 35 and 40% ethylene glycol as vitrification solution for solid surface vitrification (SSV) and in-straw vitrification (ISV), respectively. Survival rate of vitrified–warmed oocytes, evaluated on the basis of ooplasm homogeneity, oolemma integrity and zona pellucida intactness, as well as parthenogenetic blastocyst rates of vitrified–warmed oocytes were significantly higher with SSV (89.3 and 13.6%, respectively) than ISV (81.8 and 5.5%, respectively). However, they were still significantly lower than that of control oocytes (100 and 34.2%, respectively). For examining the ultrastructural changes, fresh, VS-exposed (ISV and SSV), and vitrified–warmed (ISV and SSV) oocytes were processed for transmission electron microscopy. In VS-exposed oocytes, reduction of microvilli abundance and damage of mitochondrial membrane were found only in the ISV group. In vitrified–warmed oocytes, however, it was clear that both methods of vitrification induced profound ultrastructural modifications to microvilli, mitochondria, oolemma and cortical granules as well as to the size and position of vesicles. Damaged mitochondria were, however, more abundant in ISV vitrified oocytes than in SSV vitrified oocytes, which correlated with the developmental data, showing the superiority of the SSV method. The present study demonstrated the feasibility of vitrification of in vitro-matured swamp buffalo oocytes.

Keywords: cryopreservation, matured oocyte.


   
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