猪卵母细胞体外成熟与冷冻保存的初步研究

Primary study on in vitro maturation and cryopreservation of the porcine follicular oocytes

比较从不同卵巢卵泡直径获得的卵母细胞体外成熟能力的差异 ,并研究其冷冻保存效果。结果表明 :在体外成熟培养液中添加 10 % (体积分数 ,下同 )ECS的成熟率 (72 86 % )显著高于添加 10 %NCS (6 2 2 1% ) (P <0 0 5 ) ,而添加10 %pFF则抑制卵母细胞的体外成熟。随着卵泡直径的增大 ,卵母细胞体外成熟能力逐渐增强。采用程序冷冻保存方法 ,培养成熟组的卵母细胞成活率 (35 5 9% )显著高于培养前 (2 4 6 4 % )和培养 2 4h组 (2 3 36 % ) (P <0 0 5 )。培养 2 4h(77 2 2 % )和培养成熟 (72 81% )组 ,卵母细胞解冻后的形态完整率均显著高于培养前 (5 3 2 4 % ) (P <0 0 5 )。

Factors affecting oocytes in vitro maturation(IVM)from different size of follicles and cryopreservation of oocytes in different development phases were studied.The results showed that the maturation rate of IVM was very significantly higher in the medium containing 10%ECS(72 86%)than that containing 10%NCS(62 21%)(P<0 05).Addition of 10% porcine follicular fluid(pFF)to mature medium inhibited the in vitro maturation of oocytes.Oocytes from different size of follicles had different development ability.Oocytes collected from big(>5 mm)and medium(2?5 mm)follicles had a higher maturation rate than oocytes from small(<2 mm)follicle (P<0 05).Oocytes in MⅡ stage had higher antifreezing ability than GV stage and oocytes culture for 24 h.The survival rate of oocytes after cryopreservation and thawing was 35 59%,significant higher than oocytes cryopreservation in GV stages(24 64%)and culture for 24 h(23 36%)(P<0 05).The rate of morphological normality after thawing was significantly lower in group of GV stages(53 24%)than in group of MⅡ stage(72 81%)and culture for 24 h(77 22%)(P<0 05).