食蟹猴-猪异种体细胞核移植的初步研究  

Preliminary study on long-tailed macaque(Macaca fasicularis)-pig interspecies somatic cell nuclear transfer

作　　者：吕培茹[1,2] 刘红波[1,2] 左二伟[1,2] 杨小淦[1,2] 宁淑芳[1,2] 韩晓慧[1,2] 陆阳清[1,2] 卢晟盛[1,2] 卢克焕[1,2] 

机构地区：[1]广西大学广西亚热带生物资源保护利用重点实验室,广西南宁530004 [2]广西大学动物科技学院,广西南宁530004

出　　处：《中国兽医学报》2011年第2期258-263,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30860039);广西科学研究与技术开发计划资助项目(桂科能0815011-6-3)

摘　　要：采用食蟹猴耳部成纤维细胞作为核供体利用电融合法构建猴-猪异种核移植胚胎,研究其核重编机制并寻找适宜的培养基,初步建立食蟹猴-猪异种核移植体系。结果显示:(1)重构胚胎激活后3 h,供体核的大小基本不发生变化,但在激活后6 h大部分形成膨大的类原核。(2)食蟹猴-猪异种核移植胚胎融合率为74.2%,有70.5%的重构胚胎发生卵裂,29%的卵裂胚胎发育到桑椹胚阶段。(3)在NCSU-23+10%FCS和TCM199+10%FCS培养基中,重构胚突破4-细胞阻滞发育到8-细胞以上的比例分别为23.5%和17.0%,在NCSU-23培养基中的为19.9%,但是只有在NCSU23+10%FCS和TCM199+10%FCS培养基中重构胚胎发育到囊胚阶段(1.4%vs1.1%),尽管差异不显著。本研究首次建立了食蟹猴-猪异种体细胞克隆体系。Long-tailed macaque ear fibroblast cells,as donor cells,were introduced into pig enucleated oocytes to investigate whether they could be reprogrammed.To investigate appropriate embryo culture conditions for interspecies cloned embryos,three culture mediums were used,such as NCSU-23,NCSU-23＋10%FCS,TCM199＋10%FCS.Results showed that 6h after activation,the introduced nucleus gradually became swollen,indicating that monkey ear fibroblast cells could be reprogrammed.The fusion rate of monkey-pig reconstruct embryos was 74.2%;70.5% of reconstructed embryos cleaved and 29% cleaved embryos developed to morula;all the three culture media could help the reconstructed embryos break through 4-cells block and develop to beyond 8-cells（23.5%,17.0% vs 19.9%,P0.05）.And a few reconstructed embryos developed to blastocysts（1.4% vs 1.1%,P0.05） in media of NCSU23＋10%FCS,and TCM199＋10%FCS,but there was no significant difference between two media.In conclusion,we firstly established a system for long-tailed macaque-pig interspecies somatic cell nuclear transfer.

关 键 词：猪 食蟹猴 异种核移植 核重塑 培养基 

分 类 号：S852.1[农业科学—基础兽医学]