Zakiah Saumi, - and Viski Fitri Hendrawan, - and Widjiati, - and Epy Muhammad Luqman, - (2022) Effect of Dosage on Combination Ethylen Glycol and Dimethyl Sulfoxide ss Cryoprotectant Using Vitrification on Viability of Mice (Mus Musculus) Blastoscyst Pasca Warming. Ecology, Environment and Conservation, 28 (3). pp. 1125-1129. ISSN 0971–765X

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Abstract

Cryopreservation is a method of reproductive technology that is useful for embryo and preservation creatures that are almost extinct. Cryoprotectants are often used for vitrification are PROH, DMSO and EG because they have good and efficient results. The study objective was to know does the viability of mice blastocyst increase with the combination of intracellular cryoprotectant ethylene glycol (EG) and dimethyl sulfoxide (DMSO) in vitrification. This study used mice oocytes then carried out in vitro fertilization and cultured until reaching the blastocyst stage. The mice were superovulated using 5 IU PMSG and 5 IU hCG after 48 hours, then monomatting using infertile males. Egg cells are taken by tearing the fertilization bag and washing using MEM. Embryo is cultured so that it reaches the blastocyst phase and is then divided into 4 groups: vitrified using 30% EG, 30% DMSO, 20% combination EG + 10% DMSO and 10% P4 combination EG and 20% DMSO. The four groups were inserted into the hemi straw which has been modified with 0.25 berurn and then put back into the hemi straw with a size of 0.5. Hemi Straw which already contained embryos was inserted into a liquid N2 container with a temperature of -196. Warming was done after the embryo was vitrified for a week. After being removed from the container, heating is done at room temperature. The embryo hemi straw was inserted into the sucrose level alternately, i.e. 0.25 M, 0.5 M and 1M respectively for 2 minutes. Warming process to remove cryoprotectants that are in the embryo. Examination of the viability of blastocyst was done using an inverted microscope. The results of this study that there were significant differences between groups using combination cryoprotectant EG 20% and DMSO 10% (p<0.05) with 84% viability. The combination of EG and DMSO could increase the viability of mice blastocyst. The ethylene glycol 20% and dimethyl sulfoxide 10% was the best concentration combination based on this research

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