【摘要】 本研究主要探讨水牛体细胞核移植电融合的影响因素,以摸索出适合水牛体细胞核移植的电融合条件。结果显示:直流脉冲前施加2 .0 V交流电波能显著提高Φ<1 5 μm供体细胞的融合率及胚胎的卵裂率( P<0 .0 5 ) ,但对其存活率和囊胚率以及2 0 μm<Φ<3 0 μm供体细胞的融合率、胚胎的存活率、卵裂率及随后的囊胚发育率无显著影响( P>0 .0 5 ) ;然而无论施加交流电与否,直径2 0～3 0 μm供体细胞的囊胚率均显著高于直径小于1 5 μm供体细胞的囊胚率( P<0 .0 5 )。注核操作液中添加聚乙烯醇( PVA)对重组胚的融合率和卵裂率没有显著影响( P>0 .0 5 ) ,但激活后重组胚的存活率( 95 .83 %)及囊胚发育率( 1 1 .3 2 %)显著高于对照组的重组胚存活率( 74.77%)和囊胚率( 3 .75 %,P<0 .0 5 )。结果表明:( 1 )直流脉冲前施加2 .0 V交流电波能提高直径较小供体细胞的融合效果;( 2 )注核操作液中添加0 .1 %聚乙烯醇有利于提高水牛体细胞核移植的效果更多还原

【Abstract】 Factors affecting the electrofusion of buffalo somatic cell nuclear transfer were investigated in this study.Infliction of2 .0 V alternating current( AC) before direct current( DC) pulses resulted in a significant increase in the fusion rate and cleavage rate of embryos reconstructed with donor cellsin diameter less than 1 5μm,butthe blastocystdevelopmentwas still lower than that of embryos reconstructed with donor cells in diameter of2 0～ 3 0 μm in spite of AC application. Addition of polyvinylalcohol( PVA) to manipulation medium increased the survival rate and blastocyst developmentof NT embryos.In conclusions,application of2 .0 V AC before DC pulses could improve the fusion rate of donorcellsin diameterlessthan1 5μm and addition of PVA to manipulation medium of PVA to manipulation medium improved the developmentof buffalo NT embryos.更多还原