绵羊卵丘细胞细胞周期同步化对核移植胚胎发育的影响

doi:10.11843/j.issn.1671-7236.2015.03.023

《中国畜牧兽医》---唯一指定的官方网站 ›› 2015, Vol. 42 ›› Issue (3): 658-662.doi: 10.11843/j.issn.1671-7236.2015.03.023

绵羊卵丘细胞细胞周期同步化对核移植胚胎发育的影响

苏文龙¹, 李璐¹, 孟娜娜¹, 刘梦鹤¹, 王寒阳¹, 李捷鑫¹, 曹慧¹, 田树军^1,2, 孙树春^1,2, 李俊杰^1,2,3

1. 河北农业大学动物科技学院, 保定 071000;
2. 河北省牛羊胚胎工程技术研究中心, 保定 071000;
3. 河北农业大学动物医学院, 保定 071000

收稿日期:2014-09-01 出版日期:2015-03-20 发布日期:2015-03-30
通讯作者: 李俊杰 E-mail:lijunjie816@163.com
作者简介:苏文龙(1988-),男,河南焦作人,硕士生,研究方向:动物繁殖与胚胎工程,E-mail:suwenlong198865@163.com
基金资助:
河北省自然科学基金 (C2014204119);河北省高等学校科学技术研究项目(ZD2014002)

Effects on Somatic Cell Nuclear Transfer Embryos Development of Cell Cycle Synchronization of Sheep Cumulus Cells

SU Wen-long¹, LI Lu¹, MENG Na-na¹, LIU Meng-he¹, WANG Han-yang¹, LI Jie-xin¹, CAO Hui¹, TIAN Shu-jun^1,2, SUN Shu-chun^1,2, LI Jun-jie^1,2,3

1. College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China;
2. Research Center of Cattle and Sheep Embryo Engineering Technique of Hebei, Baoding 071000, China;
3. College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China

Received:2014-09-01 Online:2015-03-20 Published:2015-03-30

摘要/Abstract

摘要： 供体细胞周期同步化是影响体细胞核移植成功率的重要因素之一.试验分别对绵羊卵丘细胞采用血清饥饿和接触抑制的方法进行细胞周期同步化处理,使用流式细胞仪检测各组细胞周期的分布.结果发现,与对照组相比,卵丘细胞经血清饥饿24~72 h后,显著地增加了G₀/G₁期细胞的百分比(P <0.05);接触抑制24~72 h,G₀/G₁期细胞所占比例与血清饥饿组无显著差异(P >0.05),但显著高于对照组(P <0.05);用经血清饥饿与接触抑制的供体细胞进行核移植后,重构胚卵裂率、桑椹胚率和囊胚率差异不显著(P >0.05),但二者囊胚率显著高于对照组(P <0.05).上述结果证实,血清饥饿和接触抑制均能使绵羊卵丘细胞周期同步化至G₀/G₁,均可用作绵羊体细胞核移植的供体细胞细胞周期同步化处理.

关键词: 绵羊; 卵丘细胞; 细胞周期同步化; 体细胞核移植

Abstract: The cell cycle synchronization of donor cell is one of the important factors on success of somatic cell nuclear transfer.In this study,we treated the sheep cumulus cells with serum starvation and contact inhibition,then analyzed cell cycle by flow cytometry.The results showed that compared to the control group,24 to 72 h serum starvation increased the proportion of cells at the G₀/G₁ phase (P <0.05).The proportion of cells at the G₀/G₁ phase was increased after contact inhibition 24 to 72 h,but there was no significnant difference between serum starvation and contact inhibition groups (P >0.05),and they were significant higher than control group (P <0.05).Then used these cumulus cells synchronized by serum starvation or contact inhibition as SCNT donor cells,there was no significant difference in the cleavage,morulae and blastocyst rates of two groups (P >0.05),and blastocyst rate of two groups was significant higher than control group (P <0.05).The results suggested that sheep cumulus were effectively synchronized at G₀/G₁ stages by serum starvation or contact inhibition,and could be used as donor cells.

Key words: sheep; cumulus cells; cell cycle synchronization; somatic cell nuclear transfer

中图分类号:

S814.8

苏文龙, 李璐, 孟娜娜, 刘梦鹤, 王寒阳, 李捷鑫, 曹慧, 田树军, 孙树春, 李俊杰. 绵羊卵丘细胞细胞周期同步化对核移植胚胎发育的影响[J]. 《中国畜牧兽医》---唯一指定的官方网站, 2015, 42(3): 658-662.

SU Wen-long, LI Lu, MENG Na-na, LIU Meng-he, WANG Han-yang, LI Jie-xin, CAO Hui, TIAN Shu-jun, SUN Shu-chun, LI Jun-jie. Effects on Somatic Cell Nuclear Transfer Embryos Development of Cell Cycle Synchronization of Sheep Cumulus Cells[J]. , 2015, 42(3): 658-662.

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绵羊卵丘细胞细胞周期同步化对核移植胚胎发育的影响

Effects on Somatic Cell Nuclear Transfer Embryos Development of Cell Cycle Synchronization of Sheep Cumulus Cells

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