摘要: |
[目的]以绵羊卵母细胞为受体,构建绵羊-绵羊和山羊-绵羊体细胞克隆胚,比较其体外发育能力,探讨绵羊卵母细胞对异种体细胞核的再程序化能力.[方法]以体外成熟的绵羊卵母细胞为核受体,分别以绵羊及山羊胎儿成纤维细胞为核供体,经卵母细胞去核、注核、重构胚电融合、激活等一系列操作,构建同种绵羊-绵羊及异种山羊-绵羊体细胞的克隆胚,比较其体外发育能力.[结果]同种间克隆胚的囊胚发育率(16.0%)高于异种间克隆胚(7.4%).异种间克隆胚在8-细胞到16-细胞及16-细胞到桑椹胚期间的发育能力显著低于同种间克隆胚.2种克隆胚在2-细胞到8-细胞期间的发育速度基本相同,但异种间克隆胚在8-细胞到囊胚期间的发育速度明显低于同种间克隆胚,两者相差12~24 h.[结论]绵羊卵母细胞对异种体细胞核具有再程序化能力,但这种再程序化能力明显低于其对同种体细胞核的再程序化能力. |
关键词: 体细胞核移植 体外发育 绵羊 山羊 |
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基金项目:国家高技术研究发展计划(863计划),内蒙古科技大学科研基金? |
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Construction sheep-sheep and goat-sheep somatic cell cloned embryos and their development in vitro |
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Abstract: |
【Objective】 Sheep-sheep and goat-sheep somatic cell cloned embryos were constructed,their developmental capacity in vitro,was compared,so as to investigate the capacity of sheep oocytes reprogramming interspecies somatic nucleus.【Method】 Using in vitro matured sheep oocytes as nuclear recipient and sheep(SFFs) or goat fetal fibroblasts (GFFs) as nuclear donor,intra sheep-sheep and interspecies goat-sheep cloned embryos were constructed after series of manipulations of enucleated oocytes,injecting donor cell,electrofusing and activating constructed embryos;in addition,their developmental capacity in vitrowere compared.【Result】 The rate of intra blastocyst (16.0%) was higher than that of interspecies blastocyst (7.4%).The developmental ability of interspecies cloned embryos was lower than that of intraspecies cloned embryos during 8-cell to 16-cell stage and 16-cell to morula stage.Moreover,during 2-cell to 8-cell stage,the developmental speeds of two cloned embryos were identical;however,during 8-cell to blastocyst stage,the developmental speed of inter was 12-24 h slower than that of intraspecies cloned embryos.【Conclusion】 Sheep oocytes have the capacity of reprogramming interspecies nucleus,but the capacity of sheep oocyte reprogramming interspecies nucleus was lower than that of sheep oocyte reprogramming intraspecies nucleus. |
Key words: somatic cell nuclear transfer development in vitro sheep goat |