卵母细胞和供核体细胞质量对牛体细胞核移植效率的影响

Effect of the quality of oocytes and donor somatic cells on the efficiency of SCNT

将未成熟牛卵母细胞进行体外成熟培养,挤压法去除卵母细胞核,以牛耳成纤维细胞作为供核细胞进行体细胞核移植研究,观察形成囊胚组与未形成囊胚组平均每卵巢获卵数、卵母细胞体外成熟率、供核体细胞细胞传代次数、饥饿天数的差异。结果表明:形成囊胚组平均每卵巢获卵数(4.90枚)和体外成熟率(63.9%)均显著高于未形成囊胚组(分别为4.05枚、48.4%)。形成囊胚组供核体细胞代数(4.7代)和血清饥饿天数(4.7d)与未形成囊胚组(分别为6.1代、4.4d)无显著差异。卵巢质量和卵母细胞质量是决定能否形成克隆囊胚的重要影响因素,在供体细胞传代3～8代、饥饿1～9d的范围内,供体细胞传代次数和血清饥饿处理时间对克隆效率无显著影响。

Bovine ears＇ fibroblast were used as donor cells,oocytes matured in vitro were used as recipient cells and bovine reconstructed embryo were obtained by somatic cell nuclear transfer（SCNT）.The oocytes number of per ovary,maturation rate of in vitro matured oocytes,the passages and starvation-induced period of donor cells between the blastula group and the non blastula group were compared.The oocytes number of per ovary and maturation rate of in vitro matured oocytes of blastula group were significantly higher than those in non blastula group.There were no significant difference between the two groups when the donor cells were 3-8 passages and starvation-induced 1-9 days.It was concluded that the quality of ovary and oocyte was important for the production of SCNT blastula.The passages of donor somatic cells and starvation-induced period had no significant influence on SCNT efficiency when the donor cells were 3-8 passages and starvation-induced 1-9 days.