有无透明带猪卵母细胞电激活参数的优化及体外培养方式的建立 |
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引用本文: | 任子利,赵彦玲,何若钢,卢晟盛,陆阳清,蓝海恩,潘天彪,黄敏瑞,卢克焕. 有无透明带猪卵母细胞电激活参数的优化及体外培养方式的建立[J]. 安徽农业科学, 2008, 36(33) |
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作者姓名: | 任子利 赵彦玲 何若钢 卢晟盛 陆阳清 蓝海恩 潘天彪 黄敏瑞 卢克焕 |
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作者单位: | 1. 广西亚热带生物资源保护利用重点实验室,广西南宁,530005;广西大学动物科学技术学院,广西南宁,530005 2. 广西大学动物科学技术学院,广西南宁,530005 3. 广西畜牧研究所,广西南宁,530001 |
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基金项目: | 国家自然科学基金资助项目 , 广西科学基金资助项目 , 广西科学研究与技术开发计划项目 |
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摘 要: | [目的]摸索无透明带猪体外成熟卵母细胞(ZFOs)电激活的条件和无透明带孤雌激活胚胎(ZFEs)的体外培养方式,为手工克隆法生产广西巴马小型猪体细胞核移植奠定基础。[方法]用不同场强、脉冲时程和脉冲次数激活ZIOs、ZFOs,用不同培养方式(微滴法和微穴法)培养ZIEs和ZFEs,检查第7天的囊胚率及囊胚的总细胞数。[结果]有带猪体外成熟的卵母细胞电激活最佳场强和脉冲时程为125 V/(mm.80μs),微滴法培养的囊胚发育率为(30.54±11.06)%;无带猪体外成熟的卵母细胞电激活最佳场强和脉冲时程为85V/(mm.80μs),微穴法培养,其囊胚发育率为(12.77±6.98)%;单次脉冲的激活效果好于多次脉冲。[结论]对有无透明带胚胎的培养方式,微穴法培养无透明带胚胎,微滴法培养有带胚胎较合适,一次脉冲可以有效激活卵母细胞。
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关 键 词: | 猪 有无带卵母细胞 微穴法 无透明带培养 电激活 |
Optimization of Electro-activation Parameter for Zona-intact and Zona-free Porcine Oocytes Matured in vitro and Establishment of in vitro Culture Method |
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Abstract: | [Objective]The aim was to explore the best conditions of electro-activation condition for zona-free porcine oocytes matured in vitro(ZFOs)and the in vitro culture method for Zona-free porcine embryos(ZFEs),in order to lay the foundation for GuangxiBama miniature porcine somatic cell nuclear transfer embryo by hand-made cloning(HMC).[Method] Under different electrical field strength and the pulse duration,ZIEs and ZFEs were cultured by different methods(embryos in drops and wells) for 7 d in vitro.Blastocyst development rate of embryos and the total cell number of blastocysts were examined.[Result] The optimum electrical field strength and the pulse duration were 125 V/(mm·80 μs),with a blastocyst development rate being(30.54±11.06)%for zona-intact activation embryo in drops.As for zona-free activation,embryos in wells under the conditions of electrical field strength and the pulse duration at 85 V/(mm·80 μs),the blastocyst development rate was(12.77±6.98)%.The activation effect of the multiple pulse was better than multiple pulses.[Conclusion]As for zona-intact and zona-free porcine embtyos in vitro(ZIEs and ZEFs),it was appropriate for ZIEs indrops and ZFEs in well.Oocytes was most effectively actived by a single pulse. |
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Keywords: | Porcine Zona-intact and zona-free porcine oocytes Well method Culture of zona-free embryos Electro-activation |
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