【摘要】 用成年绵羊耳部皮肤细胞进行培养传代。将培养出的成纤维细胞经血清饥饿法处理后 ,作为核供体移植到MⅡ期去核卵母细胞中 ,重构胚经电融合与化学激活后 ,在 38.6℃、5%CO2 、最大相对湿度条件下进行体外培养 (IVC)。用细胞松驰素B法与离子霉素 +6-DMAP法在电融合后进行化学激活。重构胚卵裂率分别为 37.8%与 4 3.8% ( 51/135与 81/185) ,囊 /桑堪胚率分别为9.8%和 12 .3% ( 5/51和 10 /81)。在融合电压 1.5Kv/cm、持续时间 30us、脉冲次数为 2次的条件下 ,孤雌激活胚胎的卵裂率和囊 /桑堪胚率分别为 81.0 %和 12 .9%。更多还原

【Abstract】 Sheep Fibroblast cells derived from adult sheep ear were cultivated and passaged. Fibroblast cells were transferred to enucleated oocytes of MII phase by micromanipulation.The donor cells used for nuclear transfer were synchronized at presumptive G\-0 stage by serum starvation.The reconstructed embryos were cultivated in 38.6℃,5%CO\-2,humitified incubator after electrofusion and chemical activation.Cytochalasin B and ionomycin along with 6-dimethylaminop urine (6-DMAP) were used as activating agent.The cleavage rate of reconstructed embryos activated by Cytochalasin B and ionomycin /6-DMAP were 37.8% and 43.8% (51/135 and 81/185),morulas/blastocysts rate were 9.8% and 12.3%(5/51 and 10/81) correspondently. Cleavage rate activated by arthenogenetically was 81.0%.The rate of morulas/blastocysts was 12.9% under the stimulation by two electric pulses of 1.5Kv/cm for 30μsec.更多还原