实验用小型猪胎儿成纤维细胞和骨髓间充质细胞为核供体生产转基因克隆胚胎

目的比较不同类型体细胞对生产转基因克隆胚胎效率的影响。方法利用脂质体介导的方法将质粒pEGFP-N1转染到五指山小型猪胎儿成纤维细胞和骨髓间充质细胞,经过G418筛选后均获得了阳性细胞株。然后分别以两种类型转基因细胞以及未转基因细胞为核供体进行体细胞核移植,比较不同类型供体细胞克隆胚胎的囊胚发育率。结果胎儿成纤维细胞和骨髓间充质细胞克隆胚的囊胚发育率差异不显著(P>0.05,8.3%vs.7.1%);转基因胎儿成纤维细胞(9.6%)和转基因骨髓间充质细胞(9.9%)克隆胚胎的囊胚发育率差异不显著(P>0.05,9.6%vs.9.9%);每一种类型供体细胞转基因与否对克隆胚的囊胚发育率无影响(P>0.05)。结论通过体细胞核移植技术,小型猪骨髓间充质细胞与胎儿成纤维细胞均可有效地生产转基因囊胚。

Production of Transgenic Cloned Embryos by Nuclear Transfer Using Fetal Fibroblast and Marrow Stromal Cells of Laboratory Miniature Pig

Objective The efficiency of production transgenic cloned embryos using the different types of somatic cells was examined.Methods Fetal fibroblast cells(FF) and marrow stromal cells(MSCs) of Wuzhishan pig(WZSP) were transfected with pEGFP-N1 plasmid DNA by lipofectin-mediated way.Clones were obtained by G418 selection.MSCs,FF,and GFP-MSCs,GFP-FF were used as nuclear donors for somatic cell nuclear transfer(SCNT).Results There were no significant differences in the development rates to the blastocyst stage of SCNT embryos from nontransgenic and transgenic cells between MSCs and FF(7.1% vs.8.3%,P>0.05;9.9% vs.9.6%,P>0.05).Blastocyst rate of reconstructed embryos with GFP transfected MSCs or FF in comparison to those derived from non-transgenic somatic cells did not decreased(P>0.05).Conclusion It was suggested that porcine transgenic blastocysts could be efficiently produced by SCNT using transgenic MSCs as well as FF.