Ghorbani Mina, Sadrkhanlou Rajabali, Nejati Vahid, Ahmadi Abbas, Tizroo Gholamreza
Department of Biology, Faculty of Science, Urmia University, Urmia, Iran;
Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran;
Vet Res Forum. 2012 Fall;3(4):245-9.
The effect of modified vitrification was assessed on cellular development capability in mouse embryos cultured in vitro. In this study, 466 embryos (from zygote to morula stages) were vitrified then thawed embryos have been incubated for in vitro farther development up to blastocyst stage. Also, vitrification and thawing procedures were the same for all experimental groups. Mouse different embryonic cleavage stages were vitrified in ethylene glycol (EG) plus dimethyl sulfoxide (DMSO) and sucrose (VS-1) and EG plus DMSO (VS-2) and thawed by directly placing the vitrified drop into sucrose solution (TS) at 37 ˚C. High recovery (72-97%) of morphologically normal embryos was evident following vitrification and thawing. Development of the vitrified morulae into blastocysts (92%) was higher (p < 0.05). The amount of zygote and 2-cell stages that achieved to blastocyst stage was very low. With progressing the embryo cleavage to morula stage, the embryos that reached to blastocyst were increased to its maximum number. We concluded that the modified vitrification procedure supported better survival of morula stage compared to other cleavage stages in mouse embryos.
评估了改良玻璃化法对体外培养的小鼠胚胎细胞发育能力的影响。在本研究中,466个胚胎(从合子到桑椹胚阶段)被玻璃化,然后解冻后的胚胎被培养至囊胚阶段以进行进一步的体外发育。此外,所有实验组的玻璃化和解冻程序相同。将处于不同胚胎分裂阶段的小鼠胚胎在乙二醇(EG)加二甲基亚砜(DMSO)和蔗糖(VS-1)以及EG加DMSO(VS-2)中进行玻璃化,并通过将玻璃化液滴直接置于37˚C的蔗糖溶液(TS)中进行解冻。玻璃化和解冻后,形态正常的胚胎具有较高的回收率(72-97%)。玻璃化桑椹胚发育成囊胚的比例(92%)更高(p < 0.05)。达到囊胚阶段的合子和2细胞阶段的胚胎数量非常少。随着胚胎分裂进展到桑椹胚阶段,发育到囊胚的胚胎数量增加到最大值。我们得出结论,与小鼠胚胎的其他分裂阶段相比,改良玻璃化程序能更好地支持桑椹胚阶段的存活。