供体细胞的传代次数和不同处理方法对异种核移植效率的影响

Effects of Passage Numbers of Donor Cell and Different Treatment on Interspecies Nuclear Transfer

实验旨在探讨供体细胞培养代数及其所处的细胞周期对异种核移植效率的影响。以经Hochest33342染色后去核的牛卵母细胞为受体,以处于不同细胞周期的不同代数的人成纤维细胞为供体,采用电融合方法构建重构胚,6-DAMP和乙醇激活重构胚,SOFaa培养液中培养。第20代细胞非饥饿组的融合率、发育率显著低于第20代饥饿组(P<0.05),极显著的低于第8代细胞的饥饿组和非饥饿组(P<0.01),而8-细胞胚胎率,囊胚率4组间无显著差异。结果表明,饥饿处理对于培养代数低的细胞来说可能是非必要因素,但对于高代次细胞则是提高核移植效率的必要步骤,饥饿处理可使核移植融合率和发育率显著的提高。

The effects of donor cell cycle and different passage on interspecies nuclear transfer were discussed in this research. Bovine oocytes stained by hochest33342 were enucleated as receptor and human fibroblast in different cell cycle and passage were used as donor for nuclear transfer. After electric fusion, these reconstructed embryos were activated by 6-DAMP ＋ ethanol and cultured in SOFaa medium. The fusion and development rates of the treatment in the 20th passaage cell without starvation （ P 〈 0.01 ） were significantly lower than that in the 20th passage with starvation （ P 〈 0.05 ）, and were significantly lower than that in the 8th passage with starvation and without starvation （P 〈0.01）. There were no significant differences in 8-cell embryo rates and morula-blastocyst rates in four treatments. The result indicates that starvation treatment may not be the essential factor for low passage donor cell and when donor cell is in high passage, starvation treatment can be the important way to improve the efficiency of interspecies nuclear transfer by increasing fusion rate and development rate significantly.