BCB和Zebularine对绵羊体细胞核移植胚胎发育效果的影响  

Effect of Brilliant Cresyl Blue Staining and Zebularine on Development of Somatic Cell Nuclear Transfer Embryos in Sheep

作　　者：曹慧[1] 李俊杰[1,2] 郝永兰 刘爱菊[1] 李文瑞[1] 魏巧莉 田树军[1,2] 孙树春[1,2] 

机构地区：[1]河北农业大学动物科技学院,河北保定071000 [2]河北省牛羊胚胎工程技术研究中心,河北保定071000

出　　处：《中国畜牧杂志》2017年第3期58-62,共5页Chinese Journal of Animal Science

基　　金：河北省自然科学基金(C2014204119);河北省高等学校科学技术研究项目(ZD2014002)

摘　　要：为探究亮甲酚蓝(BCB)染色选择的卵母细胞是否有利于体细胞核移植胚胎的体外发育,本实验将卵丘-卵母细胞复合体(COCs)放入含有BCB的PBS中染色,根据细胞质颜色可将卵母细胞分成BCB^+组和BCB^-组,并以未经BCB处理的COCs作为对照组,然后将卵母细胞进行体外成熟,统计卵母细胞的成熟率。将成熟后的卵母细胞进行体细胞核移植,其中,部分BCB^+组卵母细胞所需的供体细胞利用Zebularine处理,统计体细胞核移植的卵裂率、桑椹胚率和囊胚率。结果表明:BCB^+组卵母细胞的成熟率显著高于对照组和BCB^-组(71.15%vs 65.38%,53.52%,P<0.05);BCB^+组核移植胚胎的卵裂率(87.91%vs 56.83%)、桑椹胚率(37.41%vs 21.73%)和囊胚率(21.48%vs6.82%)均显著高于BCB^-组(P<0.05);与对照组相比,BCB^+组卵裂率(87.91%vs 83.23%)和囊胚率(21.48%vs14.89%)也显著升高(P<0.05)。BCB^+组供体细胞经Zebularine处理后,胚胎发育能力进一步提高,其中囊胚率显著高于BCB^+组、对照组和BCB^-组(29.25%vs 21.48%,14.89%,6.82%,P<0.05)。To investigate whether oocytes selecting were favorable for the development of somatic cell nuclear transfer embryos in vitro by brilliant cresyl blue（BCB） staining, the cumulus-oocyte complexes（COCs） were stained in PBS containing BCB. According to the color of cytoplasm, the oocytes were divided into two groups： BCB＋ group and BCBgroup. The COCs without BCB treatment were used as the control group, then the oocytes were maturated in vitro and the maturation rate of oocytes was counted. The matured oocytes were then subjected to somatic cell nuclear transfer. The cumulus cells treated with Zebularine were used as the donor cells of some BCB＋ oocytes. The cleavage rate, morula rate and blastocyst rate were counted. Results showed that the maturation rate of BCB＋ group was significantly higher than that of control group and BCB- group（71.15% vs 65.38%, 53.52%, P0.05）; after somatic cell nuclear transfer, the development level of cloned embryos was evaluated, there is significant difference in the cleavage rate（87.91% vs 56.83%）, morula rate（37.41% vs 21.73%） and blastocyst rate（21.48% vs 6.82%） between BCB＋ group and BCB- group（P 0.05）. Compared with the control group, the cleavage rate（87.91% vs 83.23%） and blastocyst rate（21.48% vs 14.89%） of BCB＋ group were also significantly higher（P0.05）. When donor cells were treated with Zebularine, the blastocyst rate of the BCB＋ group（Zeb＋BCB＋ group） was further increased, it was significantly higher than that of BCB＋ group, control group and BCB- group（29.25% vs 21.48%, 14.89%, 6.82%, P0.05）.

关 键 词：BCB染色 ZEBULARINE 绵羊 体细胞核移植 

分 类 号：S826.3[农业科学—畜牧学]