Do Jeong Tae, Schöler Hans R
CHA Stem Cell Institute & CHA Biotech, Pochon CHA University, Seoul, Korea.
Methods Mol Biol. 2010;636:179-90. doi: 10.1007/978-1-60761-691-7_11.
Genomic reprogramming can be accomplished by five different types of methods: nuclear transfer, cell fusion, in vitro culture, introduction of egg extract, and transduction of transcription factors. We have shown that fusion-induced reprogramming is an efficient method for reprogramming differentiated somatic cells to a pluripotential state (pluripotential reprogramming)--Oct4 gene reactivation occurs within 1-2 days postfusion of somatic cells with pluripotent stem cells. Reactivation of Oct4 can be monitored by detection of the GFP signal from the Oct4-GFP transgene of somatic cells. In the current report, we fused double transgenic (OG2/ROSA26) somatic cells with pluripotent embryonic stem (ES) cells, and demonstrated the presence of the somatic cell genome in all GFP-positive ES-like colony-forming cells, confirming their identity as the cell fusion hybrids.
核移植、细胞融合、体外培养、引入卵提取物以及转录因子转导。我们已经表明,融合诱导的重编程是将分化的体细胞重编程为多能状态(多能性重编程)的一种有效方法——在体细胞与多能干细胞融合后的1 - 2天内Oct4基因重新激活。Oct4的重新激活可通过检测来自体细胞Oct4 - GFP转基因的GFP信号来监测。在本报告中,我们将双转基因(OG2/ROSA26)体细胞与多能胚胎干细胞(ES细胞)融合,并证明在所有GFP阳性的类ES集落形成细胞中存在体细胞基因组,证实它们作为细胞融合杂种的身份。