【摘要】 以兔8-16细胞的新鲜胚胎或冷冻-解冻胚胎的卵裂球作为细胞核供体,成熟的去核卵母细胞作为细胞核受体。经显微操作,将分离的单个卵裂球移入去核卵母细胞透明带内,制成322枚组合卵。对组合卵施加一个1200伏/cm（V/cm）和80微秒（μs）的直流电脉冲,使卵裂球的细胞核导入去核卵母细胞之中。由新鲜供体胚胎或冷冻-解冻供体胚胎与去核卵母细胞构成的组合卵的融合率分別为87.2%（251/288）和73.5%（25/34）。将部分已融合的细胞核移植（Nticlear Transplantation,NT）卵置于含10%胎犊血清（FCS）的TCM199培养液中,在充以5%CO₂+空气的培养箱内,于39℃下培养20小时,新鲜供体胚胎和冷冻-解冻供体胚胎的NT卵发育至2-4细胞的分裂率分别为58.0%（47/81）和53.8%（7/13）。将53枚NT卵移入同步发情的5只受体母兔的输卵管内,结果一只母兔于妊娠笫33天经剖腹手术产出1只体重为70克雌性核移植仔兔。实验结果表明兔8-16细胞阶段的新鲜胚胎和冷冻-解冻胚胎均可在细胞核移植中用作细胞核供体。更多还原

【Abstract】 Nuclear donors were blastomeres of rabbit fresh or frozen-thawed 8-16 embryos, and nuclear recipients were enucleated oocytes. Blastomere-enucleated oocyte complex eggs were made with a micromanipulation technique. A DC pulse of 1200v/cm and 80 μs was applied to induce membrane fusion between a blastomere and an enucleated oocyte. Fusion rates of complex eggs from the fresh and frozen-thawed donor embryos were 87.2% （251/288） and 73.5% （25/34） respectively. After the nuclear transplant （NT） eggs were cultured in TCM 199 supplemented with 10% fetal calf serum in CO₂ incubater at 39℃ for 20 hours, some of which developed to 2-4 cell stage. The cleavage rates of NT eggs from fresh and frozen-thawed donor embryos were 58.0% （47/81） and 53.8% （7/13） respectively. Fifty-three NT embryos were transferred to 5 recipient rabbits. As a result, one female NT rabbit （birth body weight 70g） was born caesarean section from one recipient on the 33rd day of its pregnancy. The result showed that both fresh and frozen-thawed 8-16 cell embryos could be used as nuclear donors in rabbit nuclear transpiantation.更多还原