猪体细胞胞质注射核移植研究

Research on Porcine Somatic Nuclear Transfer by Intraplasmic Injection

首例体细胞克隆动物“多利”羊诞生以来，各种体细胞核移植动物相继成功克隆，但其克隆效率很低（低于10％）以及大都表现出不同程度的器官衰竭或发育畸形等。猪体细胞核移植在异种器官移植和克隆性治疗上具有重大意义，但其克隆效率更低（1%-2％）。利用胞质注射技术对比不同供核细胞、是否经血清饥饿和不同时间段激活对重构胚发育的影响，在其重构胚卵裂率上，试验结果表明：颗粒细胞与胎儿成纤维细胞无显著差异（43．37％，45．98％；P〉0．05）；血清饥饿与否无显著差异（45．61％。50．71％；P〉0．05）；在核移植后1-3h和3-5h激活显著高于立即激活（51．82％，56．98％。8．51％；P〈0．05）。

With the birth of ＂Dolly＂ in 1997, other SCNT animals produced subsequently in recent years, but the efficiency was comparably, low（under 10%） and cloning offspring showed abnormally developing organs et al. Although porcine Somatic nuclear transfer had huge potential the efficiency is very low （1%-2%）. In this article, intraplasmic which examined the effect of the kinds of donor cells （cumulus for xenotransplantation and cloning therapy, injection in porcine SCNT has been applied cells, fetal fibroblasts）, fetal fibroblasts cultured by serum hungry or not and activation time （0h, 1 -3h, 3-5h after SCNT） on reconstructed embryo development in order to provide some ways of improving the find porcine SCNT efficiency. The results show that the cleavage rates of different donor cells have no significant difference （43.37%, 45.98%; P〉0.05）, the group of serum hungry or not have no significant difference （45.61%, 50.71%; P〉0.05）,the groups of delaying activation are significantly higher than immediately activation（8.51%, 51.82%, 56.98%; P〈0.05）.