OPS玻璃化冷冻对小鼠四倍体胚胎体外发育的影响

为探究开放式拉长细管(OPS)玻璃化冷冻对四倍体胚胎发育的影响,本实验利用2-细胞胚胎电融合法制备四倍体胚胎,再对四倍体胚胎进行OPS玻璃化冷冻,分别观察记录二倍体胚胎、四倍体胚胎以及冷冻解冻后四倍体胚胎的发育情况。结果表明:2-细胞胚胎电融合效率为96.1%;二倍体胚胎组与电融合后四倍体胚胎组的囊胚率和孵化囊胚率差异不显著;冷冻解冻后四倍体胚胎的囊胚率(100%)与四倍体新鲜组(93.3%)差异不显著,其孵化囊胚率(72.3%)较新鲜组(64.9%)显著增高(P<0.05);四倍体冷冻解冻组的囊胚细胞数(31.96)与新鲜组(32.54)无显著差异;冷冻解冻后的四倍体早期囊胚进行体外培养时其发育速度比对照组更快。可见,冷冻对小鼠四倍体胚胎的囊胚率和囊胚细胞数均无显著影响,但孵化囊胚率显著提高,且OPS玻璃化冷冻后使四倍体胚胎的发育速度更快。

Effect of OPS Vitrification on Development of Mouse Tetraploid Embryos in vitro

In order to explore the effect of open lengthened tubule(OPS) vitrification on development of tetraploid embryos,tetraploid embryos were prepared by 2-cell embryo electrofusion method, and then the tetraploid embryos were OPS vitrification. The development of tetraploid embryos after freezing and thawing, fresh diploid embryos and fresh tetraploid embryos was observed and recorded. Results showed that the efficiency of 2-cell embryo electrofusion was 96.1%. There was no significant difference in blastocyst rate and incubation blastocyst rate between diploid embryo group and tetraploid embryo group after electrofusion. After freezing and thawing, the blastocyst rate of tetraploid embryos(100%) was not significantly different from that of the tetraploid fresh group(93.3%), and the hatching blastocyst rate(72.3%) was significantly higher than that of the fresh group(64.9%)(P<0.05). There was no significant difference in the number of cyst cells between the tetraploid group(31.96) and the fresh group(32.54). The frozen thawing tetraploid early blastocysts developed faster than control group when cultured in vitro. It can be seen that freezing had no significant effect on the blastocyst rate and the number of blastocyst cells of mouse tetraploid embryos in mice, but the incubation blastocyst rate was significantly increased, and the OPS vitrification cryopreservation accelerated the development of tetraploid embryos.