不同供核细胞、处理方法及激活时间对猪核移植重构胚发育的影响

Effects of different donor cells 、treated methods and active times on the Porcine NT embryos

在不同的外界因素下,使用胞质内体细胞核注射的方法,为进一步提高猪体细胞核移植的效率来寻找新途径。利用屠宰场卵巢采集的卵母细胞经过去核,注核及激活等操作后,研究猪颗粒细胞和胎儿成纤维细胞,血清饥饿处理法及不同的激活时间等因素对猪核移植重构胚体外发育的影响.结果表明,猪颗粒细胞和胎儿成纤维细胞用作核供体与猪卵母细胞构建重构胚,在卵裂率上无显著性差异。作为供核细胞的猪成纤维细胞,经过血清饥饿与不经过血清饥饿培养,核移植重构胚的卵裂率无显著性差异。在进行核注射后,使供核细胞与胞质受体作用适当的时间(1～6h)在进行激活,有利于重构胚的发育。提示颗粒细胞和猪胎儿成纤维细胞用作核供体与猪卵母细胞构建重构胚以及作为供核细胞的猪成纤维细胞,经过血清饥饿与不经过血清饥饿培养处理后,这二者对猪核移植重构胚发育的均无明显影响。但是使供核细胞与胞质受体作经过适当的时间(1～6h)的激活,却有利于重构胚的发育。

In the different outside factors, the nuclear injection of somatic cell cytoplast is made to improve efficiency of Porcine somatic nuclear transfer （NT） for finding new ways.Porcine oocytes from ovary in slaughter were made the operation of ejection, injection, activion in nuclear. The factors of porcine cumu- lus cells and fetal fibroblasts cells, serum starvation and different active times are studied to know the effection on the development of porcine somatic NT in vitro.The donor cells of porcine cumulus cells and fetal fibroblasts cells and porcine oocytes are combined to reconstruct embryos, there are not significant- ly different in cleavage. Porcine fetal fibroblasts cells for donor are through ei- ther serum starvation or not, there are not significantly different in cleavage. Af- ter nuclear injection, the mutual effect between donor cells and acceptors are ac- tive through 1--6h. It is good to promote the development of porcine reconstruct- ed embryos.The donor cells of porcine cumulus cells and fetal fibroblasts cells and porcine oocytes are combined to reconstruct embryos, porcine fetal fibrob- lasts cells for donor are through either serum starvation or not, there are also not significantly different in cleavage. But the mutual effect between donor cells and acceptors are active through 1--6h to promote the development of porcine reconstructed embryos.