Live-cell Imaging Analysis of Mouse Embryonic Stem Cell Derivation Processes

该项目旨在确定ES细胞的性质和来源。更具体地说，我们想知道ES细胞是如何从内细胞团细胞(在ES细胞派生过程中发生的事件)衍生出来的，以及它们来自什么类型的细胞。为了解决这些问题，我们决定制造多只报告小鼠，并进行活细胞成像分析。我们制作的其中一只报告小鼠可以可视化生活条件下的DNA甲基化状态。该报告小鼠被命名为甲基RO(ROSA26基因座的甲基化探针)，并保存在RIKEN BRC。利用甲基RO，我们发现DNA甲基化模式在胚胎发育早期经历了动态变化，而在ES细胞中DNA甲基化模式是相当静态的。我们现在正准备使用这种甲基小鼠胚胎对ES细胞衍生过程进行活细胞成像分析(Ueda等人正在准备中)。

This project aims to identify the nature and origin of ES cells. More specifically, we wanted to know how ES cells are derived from inner cell mass cells (the events that take place during ES cell derivation), and what kind of cells they originated from. To address these questions, we decided to generate multiple reporter mice and carried out live-cell imaging analysis. One of the reporter mice which we generated can visualize DNA methylation status in living condition. This reporter mouse was termed MethylRO(methylation probe in ROSA26locus) and deposited to RIKEN BRC. Using MethylRO, we discovered that DNA methylation pattern undergoes dynamic changes during early embryonic development, whereas DNA methylation pattern was fairly static in ES cells. We are now preparing to carry out live-cell imaging analysis of ES cell derivation process using this MethylROmouse embryo (Ueda et al.In preparation).