中国畜牧兽医

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猪去核无透明带卵母细胞电融合参数的探讨

张斌1,华再东2,郑新民2,许生成1   

  1. 1.青海大学农牧学院,青海西宁 810016;2.湖北省农业科学院,动物胚胎工程及分子育种湖北省重点实验室,湖北武汉 430064)
  • 收稿日期:2014-07-22 出版日期:2014-04-20 发布日期:2014-05-27
  • 通讯作者: 许生成,教授。E-mail:xsc0828@126.com
  • 作者简介:张斌(1988—),男,云南人,硕士生,研究方向:胚胎工程。
  • 基金资助:

    国家转基因生物新品种培育重大专项(2013ZX08006-002、2013ZX08010-3、2013ZX08006-003);湖北省农业科技创新中心课题(2011-620-001-003);湖北省国际科技合项目(2009BFA012)。

Exploration on Electrical Fusion Parameters of Enucleated Zona-free Oocytes

ZHANG Bin1, HUA Zai-dong2, ZHENG Xin-min2, XU Sheng-cheng1   

  1. 1. Agriculture and Animal Husbandry College of Qinghai University, Xining 810016, China;2. Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding, Hubei Academy of Agriculture Science, Wuhan 430064, China
  • Received:2014-07-22 Online:2014-04-20 Published:2014-05-27

摘要: 试验通过采用不同场强、电场时程和脉冲次数对去核的猪体外成熟卵母细胞电融合参数进行比较研究,观察细胞融合情况。结果表明,在电场时程30 μs、1次直流脉冲(DC)的情况下,电场强度0.8 kV/cm 时卵母细胞的融合率为94.7%,显著高于其他各组(P<0.05);当电场强度为0.8 kV/cm、1次直流脉冲的情况下,电场时程60和90 μs时,卵母细胞融合率为86.5%和83.8%,差异不显著(P>0.05),但显著低于30 μs时的电融合效率(P<0.05);在电场强度为0.8 kV/cm,电场时程为30 μs的情况下,1次电脉冲激活卵母细胞的融合率(94.7%)和2次电脉冲的融合率(95.6%)无显著差异(P>0.05),但两者显著高于3次电脉冲的融合率(81.7%)(P<0.05)。通过对各组进行比较,在电场强度为0.8 kV/cm、电场时程为30 μs、1次直流脉冲的电刺激可以对体外成熟的猪卵母细胞取得较好的融合效果。

关键词: 无透明带卵母细胞; 融合;

Abstract: In the present study, we investigated the effect of electrical fusion parameters on pig oocytes of enucleated zona-free by using different electrical field strength and the pulse duration and then observed cell fusion. The results showed that the comparison study of different electrical fusion parametes indicated that the rate of oocytes using one DC-pulse of 0.8 kV/cm for 30 μs (94.7%) groups were significantly higher than the other treatment groups(P<0.05). The rate of fusion of the enucleated zona-free oocytes in one DC-pulse of 0.8 kV/cm for 30, 60 and 90 μs group were 94.7%, 86.5% and 83.8%, respectively, showing that 30 μs treatment group were significant higher than those of 60 and 90 μs treatment groups(P<0.05). The rate of fusion in one, double and three DC-pulse of 0.8 kV/cm for 30 μs were 94.7%, 95.6% and 81.7%, respectively, showing that double DC-pulse treatment groups were no significant higher than those of one DC-pulse treatment groups (P>0.05), but both were significant higher than three DC-pulse treatment groups, the fusion rate of them was significant higher than three DC-pulse treatment groups(P<0.05).

Key words: zona-free oocytes; fusion; porcine