不同培养液对牛体细胞核移植胚胎发育的影响  

Effects of Different Culture Media on SCNT-Derived Bovine Embryos

作　　者：熊显荣[1,2] 王丽君[1] 王勇胜[1] 许文兵[1] 张辉[1] 华松[1] 张涌[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]西南民族大学生命科学与技术学院,成都610041

出　　处：《西北农业学报》2012年第4期11-16,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：转基因重大专项重大课题(2008ZX08007-004);西南民族大学中央高校基本科研业务费专项资金(11NZYQN33)

摘　　要：旨在为牛体细胞克隆胚胎体外培养体系的建立奠定基础。利用体细胞核移植技术生产牛重构胚,将重构胚随机分成2组,分别用mSOF和G1.5/G2.5培养,统计2种培养液对牛核移植克隆胚胎发育的影响,用TUNEL荧光检测系统检测2个培养组胚胎细胞凋亡情况,同时用Real time RT-PCR检测热应激蛋白基因Hsp70和凋亡相关基因Bax在2个培养组的相对表达水平。两组的卵裂率分别为85.6%和87.0%、囊胚率分别为22.9%和27.1%,差异都不显著(P>0.05),但G1.5/G2.5组的8-细胞形成率显著高于mSOF组(P<0.05);mSOF组凋亡率为12.1%±1.9%,显著高于G1.5/G2.5组(P<0.05),G1.5/G2.5组的Hsp70和Bax相对表达量显著低于mSOF组(P<0.05)。结果表明,G1.5/G2.5培养液更有利于牛体细胞核移植胚胎的体外发育。The present study were designed to compare the effects of two culture media（mSOF and G1.5/G2.5） on the development competence of pre-implantation embryos derived from somatic cell nuclear transfer（SCNT） in bovine,for the establishment of efficient somatic cell cloned bovine embryos in vitro culture system.Utilizing somatic cell nuclear transfer technology produced bovine embryos.After activation,the reconstructed embryos were randomly allocated to mSOF or G1.5/G2.5 for culture.The development competence of cloned embryos in these two culture systems was compared in terms of cleavage rate,8-cell rate,blastocyst formation rate and apoptosis cell number on day 7 blastocyts.At the same time,we compared the expression level of stress-related and apoptosis genes（Hsp70 and Bax） between the two culture groups.Bovine SCNT-derived embryos cultured in those two media showed no significantly difference in embryo cleavage rate and blastocyst formation rate（P0.05）,but 8-cell rate was higher in G1.5/G2.5 group than mSOF group（P0.05）;SCNT-derived embryos which were cultured in G1.5/G2.5 medium were lower in apoptosis rate than mSOF counterparts（P0.05）.Moreover,real time RT-PCR results indicated that SCNT-derived embryos culture in G1.5/G2.5 show lower expression level of stress-related and apoptosis genes（Hsp70 and Bax） than in mSOF group.Results indicated that G1.5/G2.5 was more suitable for culture of bovine SCNT-derived embryos than mSOF medium.

关 键 词：体外发育 MSOF G1.5/G2.5 培养液 

分 类 号：Q813.7[生物学—生物工程]