Sanaei Batool, Movaghar Bahar, Rezazadeh Valojerdi Mojtaba, Ebrahimi Bita, Bazrgar Masood, Hajian Mehdi, Nasr-Esfahani Mohammad H
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran.
Reprod Domest Anim. 2018 Oct;53(5):1159-1167. doi: 10.1111/rda.13221. Epub 2018 Jun 25.
This study aimed to determine the optimum concentration of trehalose in solutions used for vitrification of in vitro matured (IVM) ovine oocytes. IVM oocytes were randomly divided into four experimental (vitrified) and one control (fresh) groups. Experimental groups were treated with different concentrations (0.0, 0.25, 0.5 and 1.0 M) of trehalose. After warming, some viable oocytes were exposed to 0.25% pronase to test zona pellucida hardening, whereas the others were fertilized and cultured in vitro for 8 days to evaluate their developmental competence. Blastocysts quality was assessed by differential staining and TUNEL test. Survival and developmental rates of oocytes vitrified in the presence of 0.5 M trehalose were significantly higher than those of the other vitrified groups. Furthermore, there was a significant difference between fresh and vitrified groups in total blastocyst rate. Analysis of blastocysts quality also revealed a significant difference between the group treated with 0.5 M trehalose and other groups in terms of apoptotic index. Furthermore,zona pellucida digestion time period was longer in trehalose-free (0.0 M) group compared to other groups. In conclusion, we found that IVM ovine oocytes vitrified in solutions containing 0.5 M trehalose are fertilization-competent and are able to produce good-quality blastocysts with an apoptotic index comparable to that of the fresh oocytes. Therefore, 0.5 M may be considered the optimum concentration of trehalose to be used in solutions prepared for vitrification of oocytes.
本研究旨在确定用于体外成熟（IVM）绵羊卵母细胞玻璃化溶液中海藻糖的最佳浓度。将IVM卵母细胞随机分为四个实验（玻璃化）组和一个对照（新鲜）组。对实验组用不同浓度（0.0、0.25、0.5和1.0M）的海藻糖进行处理。解冻后，一些存活的卵母细胞用0.25%的链霉蛋白酶处理以检测透明带硬化情况，而其他卵母细胞则进行体外受精并培养8天以评估其发育能力。通过差异染色和TUNEL试验评估囊胚质量。在含有0.5M海藻糖的情况下玻璃化的卵母细胞的存活率和发育率显著高于其他玻璃化组。此外，新鲜组和玻璃化组在总囊胚率方面存在显著差异。对囊胚质量的分析还显示，在凋亡指数方面，用0.5M海藻糖处理的组与其他组之间存在显著差异。此外，与其他组相比，无海藻糖（0.0M）组的透明带消化时间段更长。总之，我们发现，在含有0.5M海藻糖的溶液中玻璃化的IVM绵羊卵母细胞具有受精能力，并且能够产生与新鲜卵母细胞凋亡指数相当的高质量囊胚。因此，0.5M可被视为用于卵母细胞玻璃化溶液中海藻糖的最佳浓度。
