Leptin和ITS对牛卵母细胞体外培养和胚胎生产的影响  

Enhancing of bovine in vitro embryo production by leptin supplementation in oocyte IVM and ITS in early embryo IVC

作　　者：梅承[1] 施振旦[1] 吴嫦丽 曹善柏 

机构地区：[1]华南农业大学动物科学学院,广东广州510642 [2]广州市华美牛奶公司,广东广州510663

出　　处：《中国兽医学报》2008年第4期475-478,共4页Chinese Journal of Veterinary Science

基　　金：广州奶牛生物工程技术研究开发中心资助项目(2003U12CA011);广东省自然科学基金团队项目(04205804)

摘　　要：利用Leptin和ITS促进体外成熟和体外培养的牛卵母细胞的发育和质量,探讨提高胚胎体外生产的质量和数量的方法和技术。试验1:体外受精胚胎的培养液:添加BSA的KSOM中添加10mL/L浓度的ITS,结果使胚胎的桑椹胚率和囊胚率显著(P<0.05)高于培养液中不加ITS的对照组(桑椹胚率:43.48%vs29.07%,囊胚率:22.83%vs11.63%),卵裂率、正常分裂率和8细胞率与对照组差异不显著(P>0.05)。试验2:在卵母细胞的体外成熟液中添加10μg/L具有生物学活性的重组鸡Leptin成熟肽融合蛋白,Leptin处理组和对照组卵母细胞经体外成熟、受精后转入加有10mL/LITS的KSOM培养液进行体外培养。试验组卵裂率和正常分裂率极显著(P<0.01)高于对照组(88.96%vs66.81%和61.11%vs29.36%),8细胞率显著(P<0.05)高于对照组(84.84%vs69.57%)。Leptin处理的卵母细胞在受精后的桑椹胚率和囊胚率与对照组差异不显著,但最后的囊胚数量较对照组增加1倍多,分别为IVF卵母细胞总数的14.8%和6.4%(P<0.01)。这说明,添加Leptin对牛卵母细胞体外成熟有促进作用,可显著提高卵母细胞受精后早期胚胎的卵裂率、正常分裂率和8细胞率;加入ITS则能提高桑椹胚率和囊胚率;而Leptin和ITS的按顺序结合使用,则能大大增加体外生产胚胎的桑椹胚和囊胚的数量,从而提高胚胎体外生产的效率。In the present study the combinations of Leptin and ITS were used to improve developmental quality and quantity of in vitro produced bovine embryo, Addition of ITS into the BSA supplemented KSOM embryo maturation medium at concentration of 10 mL/L significantly （ P 〈 0.05） increased mottrla （43,48% vs 29.07% ） and blastocyst （22,83% vs 11.63% ） rates,but had no effect on cleavage and 8-cell rates （ P 〉 0.05）. Leptin priming of oocytes was carried by maturing oocytes in the maturation medium supplemented with 10 μg/L recombinant chicken leptin. Embryos derived from such oocytes by IVF and cultured in ITS supplemented medium as above had highly significantly higher cleavage than normal cleavage （88.96 % vs 66.81%, 61.11% vs 29.36%, P 〈 0.01） ,and significantly higher 8-cell rate（84,84% vs 69,57%, P 〈 0.05）in comparison with those from conventionally matured oocytes. Although the embryos from leptin primed oocytes exhibited only marginally （ P 〉 0.05 ） higher morula and blasotcyte rates than from normally matured oocytes, the total proportion of oocytes to blastocyst was highly significant higher in the former than in the latter group （14.8 % vs 6,4%, P 〈 0.01 ）, These results demonstrated that sequential use of Leptin in oocyte IVM and ITS in embryo IVC can substantially increase the efficiency of in vitro bovine embryo production

关 键 词：牛 卵母细胞体外培养 胚胎体外生产 LEPTIN ITS 

分 类 号：S814[农业科学—畜牧学] S823.8[农业科学—畜牧兽医]