小型猪克隆胚胎体外培养阶段的优化

中国畜牧兽医

小型猪克隆胚胎体外培养阶段的优化

杨希祥¹^，2，李智方^2，3，龙川²，冯冲²，储明星²，潘登科²，李娟¹

1.南京农业大学动物科技学院，江苏南京 210095；2.中国农业科学院北京畜牧兽医研究所，北京 100193；3.广西大学动物科学技术学院，广西南宁 530004

收稿日期:2013-12-11 出版日期:2014-07-20 发布日期:2014-08-21
通讯作者: 潘登科，博士，副研究员。E-mail:pangdengke2002@163.com 李娟，博士，副教授。E-mail:juanli@njau.edu.cn
作者简介:杨希祥（1985—），男，河南人，硕士生，研究方向：动物遗传育种与繁殖。
基金资助:
中央级公益性科研院所基本科研业务费专项资金项目(2012yq-2）；转基因生物新品种培育重大专项课题（2013ZX08010006-003）。

Optimization of Miniature Porcine Cloning Embryos in vitro Stage

YANG Xi-xiang¹^，2, LI Zhi-fang²^，3, LONG Chuan², FENG Chong², CHU Ming-xing², PAN Deng-ke², LI Juan¹

1. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China；2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China；3. College of Animal Science and Technology, Guangxi University, Nanning 530004, China

Received:2013-12-11 Online:2014-07-20 Published:2014-08-21

摘要/Abstract

摘要： 本试验从卵母细胞卵丘多少和初次卵裂时间早晚2个方面进行研究，旨在改善小型猪克隆方案的体外培养环节，提高克隆效率。将卵母细胞按卵丘多少分成3组，比较卵母细胞的成熟效果，取成熟效果好的卵母细胞进行后续试验；PA和SCNT试验均按初次卵裂时间早晚分成3组，在体外培养条件下，比较胚胎的卵裂率和囊胚率。结果表明，卵丘多的卵母细胞体外成熟39～40 h和卵丘少的卵母细胞体外成熟41～42 h，比不区分卵丘多少的卵母细胞体外成熟39～42 h的成熟效果要好；初次卵裂时间发生在26 h以前的胚胎比26 h之后的胚胎在数量和质量上都明显优越，前者胚胎的卵裂率和囊胚率显著高于后者，此结果在孤雌激活(parthenogenetic，PA)胚胎和体细胞核移植(somatic cell nucleartransfer，SCNT)胚胎的体外试验中均得到验证。本研究完善了小型猪克隆方案的体外培养环节，为器官异种移植提供相关技术参考。

关键词: 小型猪; 异种移植; 初次卵裂时间; 体细胞核移植

Abstract: To improve the in vitro maturation of pig oocytes and to modify the protocols of cloning, the size of cumulus-oocyte and timing of the first zygotic cleavage were mainly studied in the present study. The oocytes were divided into three groups by the number of cumulus cells, subsequently the maturation efficiency was compared, and the matured oocytes were used for the following experiments. According to timing of the first zygotic cleavage, PA and SCNT embryos were divided into three groups to compare the rates of cleavage and blastocysts in the in vitro culturing. Our results showed that a better ability of maturation was observed in the oocytes which had more cumulus cells(39 to 40 h) and few cumulus cells(41 to 42 h) than those oocytes of which the number of cumulus cells(39 to 42 h) didn’t being distinguish. Both of PA and SCNT, the embryos cleaved in 26 h had a better developmental competence compared to the embryos cleaved after 26 h, which was evaluated by the rates of cleavage and blastocysts. Therefore, in the present study, not only the miniature porcine cloning programs was improved, but also that it provided technical reference for organ xenotransplantation.

Key words: miniature porcine; xenotransplantation; timing of the first zygotic cleavage; SCNT

杨希祥，李智方，龙川，冯冲，储明星，潘登科，李娟. 小型猪克隆胚胎体外培养阶段的优化[J]. 中国畜牧兽医.

YANG Xi-xiang, LI Zhi-fang, LONG Chuan, FENG Chong, CHU Ming-xing, PAN Deng-ke, LI Juan. Optimization of Miniature Porcine Cloning Embryos in vitro Stage[J]. China Animal Husbandry & Veterinary Medicine.

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