【摘要】 本实验对昆明白系小鼠的桑椹胚实施OPS法玻璃化冷冻,研究不同管内解冻程序对胚胎体内外发育的影响,旨在为小家畜胚胎冷冻、解冻和移植提供技术参数。采用OPS冷冻两步法(10%(体积分数)EG+10%(体积分数)DMSO溶液中平衡30 s,移入EDFS30平衡25 s)保存小鼠桑椹胚。一步法解冻是将OPS直接插到盛有0.5mol/L蔗糖的离心管中分别平衡1、2、3、4、5和6 min,解冻后经体外培养结果表明,平衡2~4 min组囊胚发育率(93.58%~97.22%)与对照组(100%)无显著差异(P>0 05);实施两步法解冻时将OPS先后插入0.5和0.25mol/L蔗糖中,体外培养结果显示,先后平衡3和2 min组囊胚发育率(100%)最佳,同对照组无显著差异(P>0.05)。将以上方法管内解冻最佳组的胚胎用OPS管作移植导管,直接移植到受体子宫角中,其妊娠产仔率(38.57%)和管外解冻组(42.86%)及新鲜对照组(51.19%)均无显著性差异(P>0.05)。证明OPS玻璃化冷冻的小鼠桑椹胚可以进行管内解冻和直接移植,为小家畜胚胎移植提供了模型。更多还原

【Abstract】 In the present research,Kunming mouse morulae was vitrified by OPS(open pulled straw) method to investigate the effect of the different in-straw dilution protocol on in vivo and in vitro developmental potential of mouse embryos.The aim of this work was to provide technical details for embryo transfer and cryopreservation/thawing in small ruminant.Embryos were vitrified by two-step OPS method(embryos were equilibrated with 10%EG and 10%DMSO for 30?s,then with EDFS30 for anther 25?s).In the one-step thawing method,the OPS was directly plunged into tube filled with 0.5?mol/L sucrose solution for 1,2,3,4,5 and 6?min to dilute.After in vitro culture,the results showed that the embryos from 2-4?min groups could develop to blastocyst stage(93.58%-97.22%) as the control(P>0.05).When embryos were thawed by two-step method,the OPS was placed into 0.25?mol/L sucrose solution followed by equilibration in 0.5?mol/Lsucrose solution.The in vitro development results showed that when OPS was equilibrated in 0.5?mol/L sucrose for 3?min,then in 0.25?mol/L sucrose for 2?min,the recovered embryos showed normal blastocyst rate(100%).Embryos were transferred to the receipts using OPS as the catheter,the fetus developmental rates(38.57%) in the best in-straw dilution group showed no significant difference(P>0.05) from the conventional group(42.86%) and the control(51.19%).The above results showed that OPS vitrified mouse morulae could be successfully diluted and transfered by in-straw method,and it will provide a model for ET in small ruminant.更多还原