EGF和β-巯基乙醇对羔羊卵母细胞体外受精和体外发育的影响

摘要

[ Objective] To optimize the in-vitro culture system of oocytes of young animals. [ Method] Using the maturation medium containing EGF and β-mercaptoethanol,the fertilization,cleavage and blastocyst of oocytes were compared between lambs and adult sheep. (Result] EGF did not increase the cleavage rate and blastocyst rate of lamb oocytes. However, β-mercaptoethanol increased the blastocyst rate of lamb oocytes but had no impact on the cleavage rate of lamb oocytes,and the lamb oocytes matured in the medium containing 100 u.mol/L β-mercaptoethanol had significantly higher blastocyst rate than the control (P < 0.05). The cleavage rate and blastocyst rate were significantly lower in the lamb oocytes than in adult sheep oocytes (P < 0.05).The lamb oocytes matured in the medium containing 100μmol/L β-mercaptoethanol had significantly higher fertilization rate than the control,and their fertilization rate was not significantly different from that of adult sheep oocytes. The polyspermy rate was significantly higher in the lambs than in the adult sheep ( P < 0. 05) when no p-mercaptoethanol was used for maturation, but the polyspermy rate was not significantly between the lambs and adult sheep when 100 μmol/L β-mercaptoethanal was used for matu ration. The non-fertilization rate was not significantly higher in the lambs without β-mercaptoethand treatment than in the adult group and lambs with β-mercapto-ethand treatmed. [Conclusion] The p-mercaptoethanol at a supplementary dosage of 100 μmol/L could improve the developmental competence of lamb oocytes,increase fertilization rate and decrease polyspermy rate,but the developmental competence of lamb oocytes was still significantly lower than that of adult sheep oocytes.%[目的]为优化幼畜卵母细胞体外培养体系提供理论基础.[方法]研究在成熟液中添加EGF和β-巯基乙醇对绵羔羊卵母细胞受精和卵裂、囊胚情况的影响,并与成年绵羊卵母细胞进行比较.[结果]在成熟液中添加不同浓度的EGF对羔羊卵母细胞卵裂率和囊胚率无显著影响(P＞0.05).添加不同浓度的β-巯基乙醇可以提高羔羊卵母细胞囊胚率,其中添加100μmol/L β-巯基乙醇有显著影响(P＜0.05),但对卵裂率影响不显著(P＞0.05).羔羊卵母细胞卵裂率和囊胚率均显著低于成年羊(P＜0.05).添加100 μmol/L的β-巯基乙醇能显著提高羔羊卵母细胞正常受精率(P＜0.05),且与成年羊卵母细胞受精率差异不显著(P＞0.05).未添加β-巯基乙醇多精入卵率显著高于成年羊(P＜0.05),而添加100 μmol/L的β-巯基乙醇多精入卵率与成年羊差异不显著(P＞0.05).成熟液中不添加β-巯基乙醇时未受精率(20.0％)高于成年羊组(12.3％)和β-巯基乙醇(13.5％),但差异不显著(P＞0.05).[结论]添加100 μmol/L的β-巯基乙醇能显著提高羔羊卵母细胞发育能力,提高正常受精的比例并减少多精子受精的发生,但羔羊卵母细胞发育能力仍显著低于成年羊.