Abstract
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.
Highlights
Some in vitro fertilization (IVF) centers in worldwide undergo forbidden regulation, such as embryo cryopreservation and gamete donation
Slow freezing as one of the oocyte cryopreservation method were performing some limitations such as low oocyte survival rate,[7, 9, 10] increased risk of oocyte ageing[7, 11], and reduced embryo development compared to the fresh cycle.[7]
At 8-cells cleavage state, this study showed significant differences in the morphological grading of Grades 1 and 5. (Fig. 3) There were significantly more abundant of Grade 1 embryos observed in Kitazato than in sucrose media (p=0.01). (Fig. 3A) In contrast, there were significantly more Grade 5 embryos observed in sucrose than in Kitazato media (p=0.03). (Fig. 3A) In addition, in comparison between Kitazato and trehalose media, there were not significant differences
Summary
Some in vitro fertilization (IVF) centers in worldwide undergo forbidden regulation, such as embryo cryopreservation and gamete donation. Oocyte selection and cryopreservation could be introduced as a solution.[1,2,3,4,5,6,7] In addition, other reasons such as. Cancer, endometriosis surgery and career are convincing the oocyte selection and cryopreservation methods to ensure fertility preservation.[8]. The female mice were superovulated by given 10 IU of gonadotropin (Gonal F, Serono Ferring, Germany), followed by ovulation trigger by given 10 IU human chorionic gonadotropin (HCG) (Pregnil, Hoddesdon Hertfortshire, United Kingdom) by intraperitoneal injection, 48 hours apart
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