Addition of ficoll and disaccharides to vitrification solutions improve in vitro viability of vitrified equine embryo.

The study explores the impact of using different solutions for the vitrification of horse embryos. The results indicated that solutions with sucrose and trehalose were more effective, leading to a higher percentage of viable and morphologically superior embryos.

Introduction

• The research focusses on analyzing the in vitro viability of equine embryos vitrified in three different solutions. Vitrification is a technique used to quickly freeze biological cells, like embryos, to preserve them. It has significant applications in reproductive medicine and biology.

Methods

• This study specifically used day 6 and 6.5 embryos, which were measured and evaluated for their morphology.
• The embryos were divided into three groups. Group 1 used 40 percent ethylene glycol in PBS, a common vitrification solution. Groups 2 and 3 used similar solutions enhanced with the addition of 18 percent Ficoll, an agent used to separate cells, with either 0.3M sucrose or 0.3M trehalose – both kinds of disaccharides.
• Each vitrified embryo was individually placed into a 0.25ml straw, which was then cooled and immersed in liquid nitrogen, a procedure characteristic of the vitrification process.
• After warming at 20 degrees Celsius for 20 seconds, the embryos were removed from the straws and placed in a solution of 0.5M sucrose in PBS, then transferred to a plain PBS solution.

Results

• The diameter of each embryo was re-measured post-vitrification. Their morphology and viability were evaluated using Propidium iodide and Hoechst 33258 dyes, which are tools used to reveal the viability of cells and their structure.
• The solution with sucrose resulted in 19.2 percent viability, whereas the solution with trehalose resulted in 26.7 percent viability.
• The embryos vitrified with the solutions containing these disaccharides showed the highest percentage of viable cells and were of better morphological quality compared to those in Group 1.

Conclusion

• This research study suggests that the addition of Ficoll and either sucrose or trehalose to the vitrification solution can improve the in vitro viability and morphological quality of equine embryos.