延迟激活对猪克隆胚胎体外、体内发育效率的影响 <sup>*</sup>

doi:10.13523/j.cb.20190403

中国生物工程杂志

2019, Vol. 39

Issue (4): 16-23 DOI: 10.13523/j.cb.20190403

研究报告

延迟激活对猪克隆胚胎体外、体内发育效率的影响 ^*

石俊松¹,罗绿花¹,周荣¹,麦然标¹,纪红美¹,余婉娴¹,吴珍芳²,蔡更元^2**(

)

1 温氏食品集团股份有限公司/国家生猪种业工程技术研究中心新兴 527400
2 华南农业大学动物科学学院广州 510642

Delayed Activation Can Improve in Vitro and in Vivo Developmental Capacity of Pig Cloned Embryos

Jun-song SHI¹,Lü-hua LUO¹,Rong ZHOU¹,Ran-biao MAI¹,Hong-mei JI¹,Wan-xian YU¹,Zhen-fang WU²,Geng-yuan CAI^2**(

)

1 National Engineering Research Center for Breeding Swine Industry, WENS Foodstuff Group Co.Ltd,Xinxing 527400,China
2 College of Animal Science, South China Agricultural University,Guangzhou 510642,China

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摘要：

为了提高猪克隆效率,获得更多的克隆猪,研究了延迟激活对猪体细胞克隆胚胎体外、体内发育的影响。研究发现,和同步融合激活方法相比,延迟激活虽然会降低克隆重构胚的融合率(P>0.05),但能够显著提高克隆胚胎的卵裂率(P<0.01)和囊胚率(P<0.05);延迟激活方法重构胚使用CB辅助激活4h,其囊胚率均极显著高于不使用CB组(P<0.01);将克隆胚胎移植到126头受体母猪后,延迟激活组受体母猪分娩率显著高于同步激活组(P<0.05),虽然在窝均总仔、窝均活仔、克隆效率方面没有显著差异,但延迟激活组显然获得了更多的克隆仔猪。以上结果说明,延迟激活方法能够提高猪克隆胚胎的体外、体内发育效率。

关键词： 延迟激活; 克隆; 猪

Abstract:

The influence of delayed activation on in vitro and in vivo porcine somatic cell clone was studied in order to enhance the efficiency of porcine clone so as to obtain more clone pigs. Related studies had shown that, compared with synchronous fusion activation, delayed activation could significantly improve the cleavage rate (P<0.01) and the blastocyst rate (P<0.05) of cloned embryos, although it might decrease the fusion rate of cloned reconstructed embryos (P> 0.05). CB was utilized to assist in the delayed activation for 4h to reconstruct embryos. The blastocyst rate of the delayed activation group was significantly higher than that of the group not using CB (P<0.01). Cloned embryos were transplanted into 126 recipient sows. It was shown that the parturition rate of recipient sows was remarkably higher in the delayed activation group than in the synchronous activation group (P<0.05). Although these two groups did not differ significantly in terms of average litter size, average live births and clone efficiency, it was obvious that more cloned piglets were obtained in the delayed activation group than in the synchronous group. The above-mentioned result demonstrates that delayed activation may enhance the in vitro and in vivo development efficiency of porcine clone embryos.

Key words: Delayed activation Cloning Pig

收稿日期: 2018-09-28 出版日期: 2019-05-08

ZTFLH:

S814.8

基金资助: * 国家转基因重大专项(2016ZX08006-003);广东省应用型科技研发专项资金资助项目(2016B02023306)

通讯作者: 蔡更元 E-mail: cgy0415@163.com

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引用本文:

石俊松,罗绿花,周荣,麦然标,纪红美,余婉娴,吴珍芳,蔡更元. 延迟激活对猪克隆胚胎体外、体内发育效率的影响 ^*[J]. 中国生物工程杂志, 2019, 39(4): 16-23.

Jun-song SHI,Lü-hua LUO,Rong ZHOU,Ran-biao MAI,Hong-mei JI,Wan-xian YU,Zhen-fang WU,Geng-yuan CAI. Delayed Activation Can Improve in Vitro and in Vivo Developmental Capacity of Pig Cloned Embryos. China Biotechnology, 2019, 39(4): 16-23.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20190403 或 https://manu60.magtech.com.cn/biotech/CN/Y2019/V39/I4/16

图1 延迟激活对重构胚融合率和体外发育的影响

图2 延迟激活对重构胚融合率和体外发育的影响

图3 CB处理对延迟激活克隆胚胎体外发育的影响

图4 克隆胚胎囊胚阶段和囊胚Hoechst 33342荧光染色

表1 激活方法对克隆猪分娩率的影响

表2 激活方法对克隆猪生产效率的影响

图5 使用延迟激活法出生的7头克隆小猪

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