Sumer Huseyin, Nicholls Craig, Liu Jun, Tat Pollyanna A, Liu Jun-Ping, Verma Paul J
Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, Melbourne, Australia.
Int J Dev Biol. 2010;54(11-12):1723-8. doi: 10.1387/ijdb.103204hs.
The ectopic expression of the key transcription factors Oct4, Sox2, c-Myc, and Klf-4 have been shown to reprogram somatic cells to a pluripotent state. In turn these induced pluripotent stem (iPS) cells, like embryonic stem (ES) cells, have been shown to be able to reprogram somatic cells by cell fusion. In this study we compare the differences and similarities between ES and iPS cells measured by somatic cell fusion to somatic cells harboring an Oct4-GFP transgene. We found that iPS cells were just as potent as ES cells at reprogramming the somatic genome as measured by Oct4-GFP reactivation. The resulting ES-somatic and iPS-somatic cell hybrids were characterized for expression of key pluripotency genes, immunostaining for Oct4, SSEA-1, and the ability to differentiate into cell types representative of the three germ layers. In addition to restoring pluripotency to the somatic genome following cell fusion, the telomere maintenance mechanisms of both the ES and iPS cells were found to be dominant in the resulting ES-somatic and iPS-somatic cell hybrids, resulting in the lengthening of the somatic telomeres following cellular reprogramming. Therefore this study supports the view that iPS cells can be virtually indistinguishable from ES cells, even with regard to their reprogramming ability.
关键转录因子Oct4、Sox2、c-Myc和Klf-4的异位表达已被证明可将体细胞重编程为多能状态。反过来,这些诱导多能干细胞(iPS细胞),就像胚胎干细胞(ES细胞)一样,已被证明能够通过细胞融合对体细胞进行重编程。在本研究中,我们比较了通过体细胞融合与携带Oct4-GFP转基因的体细胞来测量ES细胞和iPS细胞之间的异同。我们发现,通过Oct4-GFP重新激活来衡量,iPS细胞在重编程体细胞基因组方面与ES细胞一样有效。对所得的ES-体细胞和iPS-体细胞杂交细胞进行了关键多能性基因表达的表征、Oct4、SSEA-1的免疫染色以及分化为代表三个胚层的细胞类型的能力分析。除了在细胞融合后恢复体细胞基因组的多能性外,还发现ES细胞和iPS细胞的端粒维持机制在所得的ES-体细胞和iPS-体细胞杂交细胞中占主导地位,导致细胞重编程后体细胞端粒延长。因此,本研究支持这样一种观点,即iPS细胞实际上与ES细胞难以区分,甚至在它们的重编程能力方面也是如此。