TSA对BCB染色筛选绵羊卵母细胞体外培养的影响  

Effect of TSA on in vitro culture of ovine oocytes screened by BCB staining

作　　者：杨楠 汪立芹[2] 林嘉鹏[2] 唐淑红[2] 吴阳升[2] 张春香[3] 黄俊成[2] 

机构地区：[1]兰州军区乌鲁木齐军区总医院妇产科,乌鲁木齐830000 [2]农业部草食家畜繁育生物技术重点开放实验室,乌鲁木齐830000 [3]山西农业大学动物科技学院,山西太谷030801

出　　处：《南方农业学报》2015年第6期1101-1105,共5页Journal of Southern Agriculture

基　　金：国家转基因重大专项项目(2011ZX08008-003);新疆科技支疆项目(201291147);新疆科技计划项目(201111113)

摘　　要：【目的】探讨曲古抑菌素A(TSA)对经亮甲酚蓝(BCB)染色分组绵羊卵母细胞体外发育的影响,为绵羊胚胎的产业化生产提供参考依据。【方法】将体外成熟的绵羊卵母细胞置于26μmol/L BCB染色液中避光染色90 min后,分别对BCB+和BCB-卵母细胞进行孤雌激活及体外受精,将获得的孤雌胚胎与体外受精胚胎置于含300 nmol/L TSA的胚胎培养液中培养10 h,再转移至未添加TSA的胚胎培养液中继续培养,连续观察胚胎的体外发育情况。【结果】在孤雌激活中,BCB+卵母细胞的卵裂率、囊胚发育率和囊胚细胞数均显著高于BCB-组(P<0.05,下同);在体外受精中,BCB+卵母细胞囊胚发育率和囊胚细胞数均显著高于BCB-组。将绵羊卵母细胞的孤雌胚胎与体外受精胚胎分别置于含300nmol/L TSA胚胎培养液中培养10 h,结果显示,BCB-卵母细胞孤雌胚胎和体外受精胚胎的囊胚发育率分别由23.33%和19.36%提升至35.65%和29.46%,但未显著影响BCB+卵母细胞的体外发育能力(P>0.05)。【结论】在胚胎培养液中添加300 nmol/L TSA(培养10 h)能显著提高BCB-卵母细胞体外胚胎的后期发育能力。【Objective】The present experiment was conducted to investgate effect of Trichostatin A（TSA） on in vitro development of ovine oocytes screened by BCB staining, in order to provide references for industry development of ovine embryo in vitro production. 【 Method 】 The ovine oocytes were stained with 26 μmol / L BCB for 90 min, Then, the oocyte with blue cytoplasmic was marked with BCB＋, oocyte with nonstaining cytoplasmic was marked with BCB-. Using BCB＋and BCB-oocytes as materials, the parthenogenetic activation and in vitro fertilization were conducted, respectively to obtain parthenogenetic embryo and vitro fertilization-embryo. These embryos were cultured in embryo culture media with 300 nmol/L TSA for 10 h, then transferred to embryo culture media without 300 nmol/L TSA. Finally, in vitro development of embryo were observed continuously. 【 Result 】 The results of parthenogenetic activation showed that, the blastocyst rate of BCB＋oocyte was significantly higher than that of control oocyte and that of BCB-oocyte（ P〈0. 05,the same below）, and the cleavage rate and blastocyst cell number of BCB-oocyte was significantly lower than that of BCB＋oocyte. The results of in vitro fertilization showed that, the blastocyst rate and blastocyst cell number of BCB＋oocyte were significantly higher than that of control oocyte and that of BCB-oocyte（P〈0.05）, The parthenogenetic embryos and fertilized embryos of BCB- oocytes were cultured in embryo culture media without 300 nmol/L TSA for 10 h, the results showed that the blastocyst rates of BCB-oocyte＇s parthenogenetic embryo and fertilized embryo significantly increased from23.33% and 19.36% to 35.65% and 29.46%, respectively, however, in vitro development of BCB＋oocyte wasn＇t be influenced（P〈0. 05）. 【Conclusion】The in vitro development BCB-oocyte＇s embryo is improved by adding 300 nmol/L TSA into embryo culture medium and culturing for 10 h.

关 键 词：绵羊 TSA BCB 卵母细胞 体外培养 

分 类 号：S826.35[农业科学—畜牧学]