Imaging pluripotency: Time-lapse analysis of mouse embryonic stem cells

Abstract

The current view of the pluripotent state is that of a transient, dynamic state, maintained by the balance between opposing cues. Understanding how this dynamic state is established in pluripotent cells and how it relates to gene expression is essential to obtain a more detailed description of the pluripotent state. In this chapter, we describe how to study the dynamic expression of a core pluripotency gene regulator— Nanog—by exploiting single-cell time-lapse imaging of a reporter mESC line grown in different cell culture media. We further describe an automated image analysis method and discuss how to extract information from the generated quantitative time-course data.