Effect of Three Different Cryoprotectant Solutions in Solid Surface Vitrification (SSV) Techniques on the Development Rate of Vitrified Pronuclear-Stage Mouse Embryos


Abstract: The objective of this study was to compare the effects of three different vitrification solutions on the development of pronuclear-stage (PN) Dinnyes mouse embryos into blastocyst stage after vitrification by solid surface vitrification (SSV) technique. For this aim, it was compared three experimental groups and a control group. Experimental groups were distinguished each other by the vitrification solution used in SSV. It was used 4 % Ethylene Glycol (EG) at 37 ºC equilibration temperature in the first group (SSV-EG), 4% Dimethyl Sulfoxide (DMSO) at room temperature in the second group (SSV-DMSO) and 4% Propylene Glycol (PG) at room temperature in the third group (SSV-PG). After vitrification, the survived embryos were cultured to blastocyst stage in KSOM. It was determined a significant difference between the groups of SSV-EG and SSV-PG at developing rate to 2-cell stage (P < 0.05). Similarly, SSV-PG demonstrated significant differences with SSV-DMSO and the control group at developing rate to 3-8-cell stage (P < 0.05). When compared the rates of developing to morula stage among the groups, it was determined significant differences between SSV-PG and the control group at (P < 0.05); and between SSV-DMSO and SSV-PG (P < 0.01). Finally, it was compared the developing rates into blastocyst stage and found that SSV-EG demonstrated significant differences with SSV-PG and SSV-DMSO (P < 0.01). This study has shown that EG, DMSO and PG with trehalose can be used effectively as a cryoprotective agent in the quick freezing of pronuclear-stage mouse embryos. Finally, additional studies are needed to optimize the SSV method in further stage mouse embryos.

Keywords: Vitrification, pronuclear-stage mouse embryos, ethylene glycol, dimethyl sulfoxide, propilen glycol, trehalose

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