摘要
本实验旨在探讨昆明白小鼠桑椹胚超低温冷冻后Oct-4表达的变化与胚胎发育潜力的关系。胚胎采用OPS法冷冻,即胚胎于10%EG+10%DMSO溶液中预处理30 s,然后再移入玻璃化溶液(EDFS30)中处理25 s,以OPS为承载器投入液氮中。毒性组胚胎未投入液氮,其他过程与冷冻组相同,新鲜胚胎为对照组。胚胎解冻后,检测Oct-4 mRNA与蛋白的表达,通过观察其形态正常率、囊胚发育率和囊胚细胞数来综合判断其体外发育能力。结果表明:冷冻组和毒性组较桑椹胚中Oct-4 mRNA的表达量对照组相比显著降低(P<0.05),蛋白表达量3组间无显著差异。桑椹胚处理后的形态正常率以及桑椹胚培养48 h后的囊胚发育率(96.67%～100%)和囊胚细胞数(89.67～92.33)3组间无显著差异。结果显示,玻璃化冷冻保存小鼠桑椹胚后多能性基因Oct-4 mRNA表达的变化并未影响其体外发育潜力。
This study was conducted to investigate the effect of cryopreservaion on oct-4 expression and the in vitro development of Kunming mouse morula. The morulas were randomly allocated into three groups： untreated （control）, exposed to vitrification solution without being plunged into liquid nitrogen （toxicity）, or vitrified by open-pulled straw method （vitrification）. After vitrification and warming, the embryos were morphologically evaluated and assessed by their development to blastocysts, or by the average cell number of blastocyst. Oct-4 expression in morula was assessed by qRT-PCR and western blotting. The results showed that the oct-4 mRNA in toxicity and vitrification groups were significantly lower （P〈0.05） than that of control. However, there was no significant difference in oct-4 expression at protein level among three groups. The blastocyst rate （96.67% to 100.00%） and the average cell number of blastocyst （89.67 to 92.33） were similar among them. The data demonstrated that the decreased oct-4 mRNA due to cryopreservation didn＇ tnegatively affect the in vitro developmental potential of mouse morula.
出处
《中国畜牧杂志》
CAS
北大核心
2013年第9期19-22,共4页
Chinese Journal of Animal Science
基金
四川农业大学"双支计划"(34270000)
大学生科研兴趣培养计划(04050895)
