作　　者： ; ; ; ; ; ; ; ;

机构地区： 玉林师范学院

出　　处： 《中国兽医科学》 2011年第5期530-535,共6页

摘　　要： 通过采集猪卵巢表面的卵母细胞,经过体外成熟和去核,供体细胞经过分离、培养和传代,注核并构建重构胚胎,融合和激活重构胚胎,以探讨供体细胞的形态、直径和传代次数对猪体细胞核移植重构胚胎的发育能力和核移植效果的影响。结果显示,表面光滑的卵丘细胞作为供体细胞核移植后的分裂率和囊胚率均显著高于表面粗糙的卵丘细胞(P<0.05),虽然融合率差异不显著(P>0.05)。直径小于15μm的供体细胞核移植后的融合率显著低于直径大于30μm的供体细胞(P<0.05)。直径20～30μm供体细胞核移植后的分裂率与直径小于15μm的供体细胞组的分裂率差异显著(P<0.05),与直径大于30μm的供体细胞组差异不显著(P>0.05)。直径20～30μm的供体细胞核移植后的分裂率与囊胚率(63.73%和21.54%)较高,囊胚率有显著差异(P<0.05)。对不同代数胎儿成纤维细胞核移植效果的研究发现,6～9代成纤维细胞的融合率显著高于3～5代和≥10代的成纤维细胞(P<0.05),但卵裂率和囊胚率差异不显著(P>0.05)。结果表明,表面光滑的供体细胞有利于重构胚胎的发育,直径20～30μm的供体细胞较适合进行核移植,用传代培养6～9代的猪胎儿成纤维细胞进行核移植较为合适。 Oocytes on the surface of porcine ovaries were collected,in vitro matured and enucleated. Donor cells were dissected, cultured and passaged. The donor nucleus was transfered into the enucleated oocyte to reconstruct an embryo. The reconstructed embryos were fused and activated in order to discuss the developmental potential of the reconstructed embryos in the morphology and diameter and passage of donor cells and explore the transfer effect in the porcine somatic cells nuclear transfer. In result, the cleav- age rate and blastocyst rate of reconstructed embryos using smooth granulosa cells as donor cells were higher than those of the rough granulosa cells as donor cells,and it were significantly different（P〈0.05）. Fusion rates in two groups were no significantly different（P〈0.05）. The fusion rate of reconstructed em- bryos using diameter 〈15μm donor cells was lower,and it was significantly different compared with the fusion rate of the diameter ）30 μm group（P〈0.05）. The cleavage rate of the diameter of 20 to 30 μm group was significant compared with the one of the diameter 〈15μm group （P〈0.05） ,but not significant compared with the diameter 〉30μm group（P〉0.05）. The cleavage rate and blastocyst rate（63.73% ＆5. 21.54%）of reconstructed embryos using diameter of 20 to 30 μm as donor cells were higher than other groups,and the blastocyst rate was significantly different（P〈0.05）. The efficiency of nuclear transfer in different culture passages of fetal fibroblast after serum starvation was examined. The fusion rate of 6 to 9 culture passages was higher than those of 3 to 5 culture passages and over 10 culture passages, but the cleavage rate and blastocyst rate among the three groups were not significantly different（P〉0.05）. In con clusion, the smooth granulose cells as donor cells were beneficial for the development of the constructed embryos,donor cells of the diameter from 20 to 30μm were better fit for porcine somatic cell nuclear transfer,and fetal fibr

关 键 词： 猪卵母细胞 体细胞核移植 供体 重构胚 效果

领　　域： [农业科学] [农业科学]