Chen Xiaopan, Wu Rongrong, Feng Shumei, Gu Bin, Dai Licheng, Zhang Ming, Zhao Xiaoli
College of Life Sciences, Zhejiang University, No. 338, Yu-Hang-Tang Road, Hangzhou, Zhejiang 310058, China.
Biochem Biophys Res Commun. 2007 Oct 19;362(2):467-73. doi: 10.1016/j.bbrc.2007.08.008. Epub 2007 Aug 9.
Embryonic stem cells (ESCs) often display high rates of apoptosis and spontaneous differentiation in routine culture, thus bring the proliferation of these cells highly inefficient. Moreover, little is known about the factors that are indispensable for sustaining self-renewal. To surmount these issues, we established transgenic mES cell lines expressing the enhanced green fluorescent protein (EGFP) under the control of the Rex1 promoter which is a key regulator of pluripotency in ES cells. In addition, we provided a simplified and improved protocol to derive transgenic mESCs from single cell. Finally, we showed that embryoid body (EB) development was faster than adherent differentiation in terms of differentiation ratio by real-time tracking of the EGFP expression. Therefore, these cell lines can be tracked and selected both in vitro and in vivo and should be invaluable for studying the factors that are indispensable for maintaining pluripotency.
胚胎干细胞（ESCs）在常规培养中常表现出高凋亡率和自发分化，从而导致这些细胞的增殖效率极低。此外，对于维持自我更新所必需的因素知之甚少。为克服这些问题，我们建立了在Rex1启动子控制下表达增强型绿色荧光蛋白（EGFP）的转基因小鼠胚胎干细胞（mES）系，Rex1启动子是ES细胞多能性的关键调节因子。此外，我们提供了一种简化且改进的方案，用于从单细胞中获得转基因mESCs。最后，通过实时追踪EGFP表达，我们发现就分化率而言，胚状体（EB）发育比贴壁分化更快。因此，这些细胞系在体外和体内均可被追踪和筛选，对于研究维持多能性所必需的因素应具有极高价值。
