Numerous studies reported that vitrification, an ultra-rapid cooling technique, seems to be highly effective and could increase oocyte survival rate rather than slow freezing. The successful of oocyte vitrification depends on the proper combination of type and concentration of cryoprotectant. This study was addressed to determine the effects of the combination of type and concentration of cryoprotectants of vitrification media, notably in the embryo development. This experimental research was conducted by using oocyte obtained from thirty-two adult female Deutschland, Denken and Yoken (DDY) mice (7-8 weeks old). The MII mice oocytes were vitrified within 24 h after retrieval using the Cryotop method with cryoprotectants as follow : sucrose (16.5% EG, 16.5% DMSO, 0.5 mol/l sucrose), trehalose (16.5% EG, 16.5% DMSO, 0.5 mol/l trehalose) and Kitazato. The embryo development and morphological grading was observed at 2-cell and 8-cells under reverse phase light microscope and inverted microscope. This study demonstrated a good embryo development and morphological grading in sucrose and trehalose vitrification media. In embryo development, trehalose medium seems more superior compared to sucrose medium, even though Kitazato was the most superior compared to both. In the morphological grading, in 2-cells embryo, there were no significant differences between the three cryoprotectants, While, in 8-cells embryo, trehalose medium appeared to be superior compared to sucrose medium, even though seemed more inferior compared to Kitazato. The appropriate type and concentration of sugar as extracellular cryoprotectant was trehalose in oocyte vitrification based on embryo development, compared to sucrose.

Introduction: One of the important factors in Assisted Reproductive Technology (ART) is embryo quality that depends on oocyte quality. Maternal nutrition in form of soybean supplementation is thought to have benefits in oocyte quality. To determine the effect soybean supplementation to the embryo quality from oocyte side. Methods: This experimental study involved female mice from Swiss strain aged 6 weeks that were divided into two groups, group with soybean (soybean group/SG) and group without soybeans (pellet group/PG). Follicles were collected and denuded to get oocytes. The oocytes were stained with MitoTracker for assessing the mitochondrial membrane potential and TUNEL for assessing the apoptotic level. Colour intensity was assessed using a confocal microscope and determined using ImageJ software. Sperms were extracted surgically from the epididymis-vas deferens and performed preparation prior to intra cytoplasmic sperm injection (ICSI) procedure. Then, the embryos were cultured and observed for the quality. Result: In TUNEL test, the average colour intensity of the SG was lower compared to the PG, significantly (p=0.03). While in the Mito Tracker test, the average colour intensity for the SG was higher compared to PG, not significantly (p=1.08). In addition, the embryo development on Day-1 and Day-3 showed more good embryo quality of SG compared to PG, significantly (p=0.03). Conclusion: Soybean supplementation improved embryo quality at the cleavage stage by decreasing apoptosis of the oocytes rather than increasing the viability the oocytes.