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目的比较OPS法玻璃化冷冻小鼠不同成熟时期卵母细胞解冻后体外成熟、体外受精及胚胎发育能力。方法收集ICR小鼠生殖泡期(GV)未成熟卵和成熟卵,分五组行OPS(open pulled straw,开放式拉细麦管)法玻璃化冷冻,Ⅰ组:直接冷冻GV期卵;Ⅱ组:GV期卵在体外培养8h后冷冻;Ⅲ组:GV期卵在体外培养16h后冷冻;Ⅳ组:GV期卵体外培养成熟(IVM)后冷冻;Ⅴ组:直接冷冻体内成熟卵。解冻后比较各组卵受精率、卵裂率及8细胞胚胎形成率。结果Ⅰ组(GV期卵)冷冻解冻后存活率达57.5%,显著高于Ⅱ组(34.1%),Ⅲ组(32.0%),Ⅳ组(29.6%)及Ⅴ组(31.0%)(P<0.01)。各组体外受精率、卵裂率及8细胞胚胎形成率比较,差异不显著(P>0.05)。结论采用OPS法玻璃化冷冻小鼠卵母细胞,冷冻GV期卵较冷冻其它成熟时期卵效果好。
OBJECTIVE: To compare the in vitro maturation, in vitro fertilization and embryo development ability of oocytes after ozing at different maturation stages in OPS vitrified mice. Methods The immature and mature eggs of gavage (GV) in ICR mice were collected and divided into five groups: vitrified OPS (open drawn straw) method. Group Ⅰ: frozen GV oocytes directly; Group: GV stage eggs were frozen in vitro for 8h; Group Ⅲ: GV stage eggs were frozen after 16h culture in vitro; Group IV: GV stage oocytes were frozen and cultured in vitro (IVM); Group V: frozen in vivo. After thawing, the egg fertilization rate, cleavage rate and 8-cell embryo formation rate were compared. Results The survival rate of group I (GV ova) frozen-thawed reached 57.5%, significantly higher than that of group II (34.1%), group III (32.0%), group IV (29.6%) and group V (31.0% 0.01). The in vitro fertilization rate, cleavage rate and 8-cell embryo formation rate in each group were not significantly different (P> 0.05). Conclusion Ovovarieal oocytes were vitrified by OPS, and the frozen GV oocytes were better than the frozen oocytes in other mature stages.