不同培养体系对猪卵母细胞体外发育潜力的影响

Effects of different culture systems on porcine oocyte developmental competence in vitro

目的:研究猪卵母细胞体外成熟的最佳条件,为用体细胞核移植的方法生产转基因克隆猪提供大量优质的MⅡ期卵母细胞。方法:采用猪卵母细胞体外成熟技术,研究了不同激素组合、猪卵泡液、半胱氨酸和颗粒细胞对猪卵母细胞体外成熟和发育潜力的影响。结果与结论:以15U/mlPMSG+20U/mlhCG成熟率最高,为(70.0±3.30)%,显著高于15U/mlFSH+30U/mlLH、10U/mlPMSG+10U/mlhCG〔成熟率分别为(45.6±5.10)%,(60.0±3.30)%〕,添加激素能促进卵母细胞的分裂,添加15%的猪卵泡液成熟率最高,为(86.7±3.35)%,显著高于5%,10%和20%组〔成熟率分别为(73.3±3.35)%,(71.2±5.08)%和(70.0±6.70)%〕,添加猪卵泡液促进了激活卵母细胞的分裂,在培养基中添加半胱氨酸和颗粒细胞,实验组和对照组成熟率差异不显著(P>0.05),但孤雌激活后卵裂率显著提高(P<0.05)。

Objective: To study the optimal condition for porcine oocyte maturation in vitro and supply abundant high quality MⅡ porcine oocyte for producing transgenic cloned pig by somatic nuclear transfer. Methods:Effects of dose and combination of hormones, PFF, cysteine and granular cell on porcine oocyte maturation in vitro and developmental competence were studied by technology of porcine oocyte maturation in vitro. Results and Conclusion: The maturation rate in medium of 15 U/ml PMSG+20 U/ml hCG was(70.0±3.30)%, higher than that in media of 15 U/ml FSH+30 U/ml LH and 10 U/ml PMSG+10 U/ml hCG〔(45.6±5.10)% and (60.0±3.30)% ,respectively〕.The medium with hormone could promote cleavage of oocytes. The maturation rate in medium of 15% PFF was (86.7±3.35)%, higher than that in media of 5%,10% and 15% PFF group〔(73.3±3.35)%,(71.2±5.08)% and (70.0±6.70)%, respectively)〕.The cleavage was promoted in the medium with PFF. The cleavage of oocytes after parthenogenetic activation was higher than controls, although there was no difference in maturation rate.