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

该项目旨在确定ES细胞的性质和起源。更具体地说，我们想知道ES细胞是如何从内细胞团细胞（ES细胞衍生过程中发生的事件）衍生而来的，以及它们起源于什么样的细胞。为了解决这些问题，我们决定产生多个报告小鼠并进行活细胞成像分析。其中一只报告小鼠可以在活体条件下观察DNA甲基化状态。该报告小鼠被命名为MethylRO（ROSA 26基因座中的甲基化探针），并保藏于RIKEN BRC。使用MethylRO，我们发现DNA甲基化模式在早期胚胎发育过程中经历动态变化，而ES细胞中的DNA甲基化模式相当稳定。我们现在正准备使用该MethylRO小鼠胚胎进行ES细胞衍生过程的活细胞成像分析（上田等，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).