College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, Peoples Republic of China.
Cell Biol Int. 2011 Aug;35(8):789-92. doi: 10.1042/CBI20100811.
The aims of this study were to determine whether stem cells could be isolated from amniotic fluid of goat fetus at terminal gestational age and to determine if these stem cells could differentiate into adipogenic cells and be transfected with a reporter gene, EGFP (enhanced green fluorescent protein). The stem cells were isolated from amniotic fluid of goat fetus at terminal gestational age, induced to differentiate into adipogenic cells in vitro and transfected with the EGFP gene using lipofection. Markers associated with undifferentiated AFS (amniotic fluid-derived stem) cells were tested by RT (reverse transcription)-PCR. The results demonstrated that AFS cells could be isolated from amniotic fluid of goat fetus at terminal gestational age and could differentiate into adipogenic cells. The EGFP gene was transfected into AFS cells successfully. EGFP gene transfection efficiency of the three groups of transgenic AFS cells were 26.0, 29.9 and 30.5%, respectively. Both transgenic and wild-type AFS cells could express Hes1 (hairy and enhancer of split 1), Oct4 (octamer-binding protein 4) and Nanog.
本研究旨在从妊娠末期山羊胎儿的羊水分离干细胞,并确定这些干细胞是否能分化为脂肪细胞,并转染报告基因 EGFP(增强型绿色荧光蛋白)。从妊娠末期山羊胎儿的羊水中分离干细胞,体外诱导分化为脂肪细胞,并用脂质体转染 EGFP 基因。通过 RT-PCR(逆转录聚合酶链反应)检测与未分化 AFS(羊水来源的干细胞)细胞相关的标志物。结果表明,可从妊娠末期山羊胎儿的羊水中分离出 AFS 细胞,并能分化为脂肪细胞。EGFP 基因成功转染到 AFS 细胞中。三组转基因 AFS 细胞的 EGFP 基因转染效率分别为 26.0%、29.9%和 30.5%。转染和野生型 AFS 细胞均可表达 Hes1(毛发和分裂增强子 1)、Oct4(八聚体结合蛋白 4)和 Nanog。