【摘要】 分离培养了成年猪前脂肪原代细胞,并对前脂肪细胞向成熟脂肪细胞进行诱导分化,油红O染色法鉴定了脂肪细胞。利用脂质体介导的方法将质粒pEGFP-N1转染前脂肪细胞,经过G418筛选获得了阳性细胞株。然后分别以前脂肪细胞、转绿色荧光蛋白(GFP)前脂肪细胞及胎儿成纤维细胞为核供体进行体细胞核移植,结果表明:前脂肪细胞(8.8%)和胎儿成纤维细胞(8.3%)的囊胚发育率无显差异著(P>0.05);与前脂肪细胞相比,转基因前脂肪细胞的囊胚发育率降低,但差异不显著(3.7%vs.8.8%,P>0.05)。前脂肪细胞为核供体生产转基因克隆胚胎,能够获得转基因囊胚。虽然囊胚发育率不高,但为生产脂肪细胞转基因克隆猪奠定了基础。更多还原

【Abstract】 Primary culture of porcine preadipocytes was established using fat tissue from an adult pig.The preadipocytes to re-differentiate induction into mature adipocytes was confirmed by oil red O staining.A porcine preadipocyte(PreA) lines were transfected with pEGFP-N1 plasmid DNA by lipofectin-mediated way.GFP-PreA clones were obtained by G418 selection.PreA,fetal fibroblast(FF),and GFP-PreA were used as nuclear donors for somatic cell nuclear transfer in pig.The results were as follows: For the blastocyst rate of cloned embryos,no significant difference was observed between PreA and FF(8.8% vs 8.3%,P > 0.05).Blastocyst rate of reconstructed with GFP transfected PreA decreased in comparison to those derived from non-transgenic PreA.(3.7% vs.8.8%,P > 0.05).The results indicate that GFP transfected PreA can produce procine transgenic blastocytes.In spite of low blastocyst rate,cloning pigs from preadipocyte is possible.更多还原