Abstract
The feasibility of cryopreservation by vitrification of Persian sturgeon (Acipenser persicus) embryos at 48 h post-fertilization stage was investigated. Vitrification is considered the most promising option. Many factors are involved in the success of the process. The choice of a proper vitrificant solutions and temperature for thawing, were the parameters considered in the present study. Six vitrificant solutions (V1-V6) were tested using a stepwise incorporation protocol. The tested solutions contained acetamide as the main cryoprotectant +3 other permeable cryoprotectants +3 non-permeable cryoprotectants. Before loading the embryos into tubes, toxicity tested was affected with these solutions. The hatching rate of embryos that had been exposed to the vitrificant solutions was analyzed and the highest hatching rate was obtained with exposure to V1. After thawing (water bath, 0 or 20°C), embryos were incubated until hatched. The highest survival rate (69.69%) was observed in samples frozen with V1 and thawed at 20°C. These results establish that cryopreservation of Persian sturgeons embryos by vitrification is possible.
Highlights
Vitrification is the solidification of a liquid brought about not by crystallization but by an extreme elevation in viscosity during rapid cooling
Our previous studies on cryoprotectants toxicity were taken into account in the design of a stepwise protocol of cryoprotectant incorporation for the vitrification of Persian sturgeon embryos [4]
Vitrificant solution V1 was considered to be suitable for embryo vitrification, but V3 not
Summary
Vitrification is the solidification of a liquid brought about not by crystallization but by an extreme elevation in viscosity during rapid cooling. Vitrification has been reported as the most promising option to cryopreserve fish embryos [1]. There is one report of flounder (Paralichthys olivaceus) embryos surviving cryopreservation by vitrification in liquid nitrogen [2], but results have not been reproducible. Our previous studies on cryoprotectants toxicity were taken into account in the design of a stepwise protocol of cryoprotectant incorporation for the vitrification of Persian sturgeon embryos [4]. The design of a proper protocol for cryoprotectant incorporation is decisive for the success of the process, but the use of adequate freezing/thawing rates is necessary [5]. The developmental ability in frozen/thawed Persian sturgeon embryos has not yet been reported by any author. The aim of the present study was to investigate the feasibility of cryopreservation of Persian sturgeon embryos by vitrification
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