| 亮甲酚蓝筛选对广西巴马小型猪孤雌激活及克隆重构胚早期发育的影响 |
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| 引用本文： | 苏克同,卢克焕,卢晟盛.亮甲酚蓝筛选对广西巴马小型猪孤雌激活及克隆重构胚早期发育的影响[J].南方农业学报,2012,43(8):1218-1222. |
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| 作者姓名： | 苏克同 卢克焕 卢晟盛 |
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| 作者单位： | 亚热带农业生物资源保护与利用国家重点实验室/广西大学动物科技学院,南宁530005广东湛江久和医院,广东湛江524094亚热带农业生物资源保护与利用国家重点实验室/广西大学动物科技学院,南宁,530005 |
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| 基金项目： | 广西自然科学基金重点项目(2010GXNSFD013021) |
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| 摘 要： | 目的]探讨亮甲酚蓝(BCB)染色是否可以筛选出发育潜能优秀的卵母细胞.方法]猪卵丘卵母细胞复合体(COCs)在体外成熟培养前用13μmol/L BCB染色,根据卵母细胞胞质蓝色的有无分为阳性(BCB+,蓝色)和阴性组(BCB-,不着色),以在不含BCB的改良杜氏磷酸盐缓冲液(m-DPBS)中处理90 min为控制对照组,直接培育作空白对照,体外成熟培养后统计各组卵母细胞的核成熟率及孤雌激活胚胎、克隆重构胚胎(供体细胞来自广西巴马小型猪)的早期发育率.结果]BCB+组卵母细胞的核成熟率(80.8%)显著高于BCB-组(44.2%)和空白对照组(72.4%);BCB+组的孤雌囊胚形成率(32.9%)显著高于BCB-组(10.1%),但与对照组(25.8%)之间无显著差异;BCB+组显微操作核移植胚胎的囊胚发育率(14.1%)显著高于BCB-组(3.4%),但与对照组之间无显著差异;各组手工克隆胚胎发育率之间无显著差异.结论]BCB染色筛选出的卵母细胞具有较好的核成熟发育潜能,但未能显著改善广西巴马小型猪显微操作及手工克隆胚胎的早期发育.
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| 关 键 词： | 广西巴马小型猪 卵母细胞 亮甲酚蓝 核成熟 孤雌激活 显微操作 手工克隆 |
Effect of brilliant cresyl blue selection on parthenogenetic activation and early development of handmade cloning embryos of Bama minipig in Guangxi |
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| SU Ke-tong,LU Ke-huan,LU Sheng-sheng.Effect of brilliant cresyl blue selection on parthenogenetic activation and early development of handmade cloning embryos of Bama minipig in Guangxi[J].Journal of Southern Agriculture,2012,43(8):1218-1222. |
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| Authors: | SU Ke-tong LU Ke-huan LU Sheng-sheng |
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| Institution: | 1 * (1 State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources/Animal Reproduction Institute, Guangxi University, Nanning 530005, China; 2 Zhanjiang Jiuhe Hospital, Zhanjiang, Guangdong 524094, China) |
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| Abstract: | Objective]The aim of this research was to evaluate whether the oocyte with superior developmental competence could be selected via brilliant cresyl blue (BCB) staining. Method]Cumulus oocyte complexs (COCs) of Bama minipig were stained with BCB (13 μmol/L) before in vitro maturation (IVM), according to the colour of oocyte cytoplasm, COCs were divided into BCB- (colourless cytoplasm) group and BCB+ group (coloured cytoplasm), taking the COCs incubated in m-DPBS without BCB for 90 minutes before IVM was treated control group while the directly cultured IVM was treated as blank control. After IVM, oocyte nuclear maturation rate, the developmental rates of parthenogenetic activation (PA), and handmade cloning (HMC) embryos in each group of Guangxi Bama minipig were calculated. Result]Oocyte nuclear maturation rate in BCB+ group (80.8%) was significantly higher than that in BCB- group (44.2%) and control group (72.4%). The blastocyst rate of PA in BCB+ group was significantly higher than that in BCB- group (32.9% VS 10.1%), but not significantly different from the blastocyst rate of the control group (25.8%). The blastocyst rate of NT in BCB+ group was significantly higher than that in BCB- group (14.1% VS 3.4%), but not significantly different than the blastocyst rate of the control group. There was no significant difference in blastocyst rate of HMC among BCB+ group, BCB- group and the control group. Conclusion]The positive porcine COCs selected with BCB staining displayed superior developmental competence of nuclear maturation, but this method could not significantly improve the developmental rate of PA, micromanipulation, and HMC embryos at early stage. |
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| Keywords: | Guangxi Bama minipig oocyte brilliant cresyl blue nuclear maturation parthenogenetic activation micromanipulation handmade cloning |
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