EFFECTIVENESS OF INSULIN TRANSFERRIN SELENIUM SUPPLEMENTATION TO VITRIFIED MICE USING HEMI STRAW ON ZONA HARDENING, EXPRESSION OF HEAT SHOCK PROTEIN 70 AND CASPASE 3

Addition of Insulin Transferrin Selenium in vitrification medium can scavenge free radicals caused by temperatures stressors due to the freezing. Insulin Transferrin Selenium is complex protein that can stimulate cell growth, prevent cell damage due to the role of antioxidants inside, so it can maintain the viability of the embryo after thawing. Insulin Transferrin Selenium can improve the quality and viability of in-vitro blastocyst culture results. The purpose of this study is to prove the effectiveness Insulin Transferrin Selenium supplementation in vitrification medium can reduce the hardening zone of mouse embrio post thawing, proving the effectiveness of Insulin Transferrin Selenium supplementation in vitrification medium on the expression of Heat Shock Protein 70 in mouse embryos post thawing, and prove the effectiveness of Insulin Transferrin Selenium supplementation the vitrification medium can decrease the expression of caspase 3 in mouse embryos post thawing. This study covered the stages of superovulation and collection eggs, in-vitro fertilization (IVF), modification vitrification medium with supplementation of Insulin Transferrin Selenium (ITS), embryo vitrification with hemi straw, embryo culture, examination of the hardening zone of mouse embryo, examination of Heat Shock Protein 70 expression and examination of caspase 3 expression in embryos post thawing.The results showed that there were differences between the addition or supplementation of Insulin Transferrin Selenium and without addition of Insulin Transferrin Selenium in the vitrification medium to hardening zone and caspase 3, but it was no differentiation in the expression of Heat Shock Protein70. The conclusion is the supplementation of Insulin Transferrin Selenium in vitrification medium decreases the hardening zone of the mouse embryos post thawing, decreases the caspase3 expression in the mouse embryos post thawing and there is no difference in the expression of Heat Shock Protein70 in the mouse embryos post thawing.