Imaging Pluripotency: Time-Lapse Analysis of Mouse Embryonic Stem Cells

• DOI: 10.1007/7651_2015_255
• Publication Date: 2015-07-10T05:41:54Z
• Authors: Anna Pezzarossa; Ana M. V. Guedes; Domingos Henrique; Elsa Abranches
• Keywords: Homeodomain Proteins; Microscopy, Confocal; Time Factors; Cell Cycle; Optical Imaging; Cell Culture Techniques; Mouse Embryonic Stem Cells; Nanog Homeobox Protein; Flow Cytometry; Cell Line; Mice; Gene Expression Regulation; Animals; Single-Cell Analysis

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.