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

Methods in Molecular Biology
Anna PezzarossaElsa Abranches

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.

Related Concepts

Single-Cell Analysis
Flow Cytometry
Pluripotent Stem Cells
Time-Lapse Imaging
Cell Culture Techniques
Nanog Homeobox Protein
Optical Imaging
Gene Expression
Mouse Embryonic Stem Cells
Cell Cycle

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