utilizing advanced flow cytometry and live-cell imaging to evaluate iPSC pluripotency During Cell Line Selection

iPSC Pluripotency During Cell Line Selection and Differentiation

Cytometry to Evaluate iPSC Pluripotency During Cell Line Selection and Differentiation

Authors: Kirsty McBain, Daryl Cole, Nina Senutovitch, Nicola Bevan and Clare Szybut | Last updated: July 2022

When generating induced pluripotent stem cells (iPSCs) for use in downstream applications, it is commonplace to perform initial checks for key attributes such as viability and pluripotency. This allows for assessment of the quality of the produced iPSC culture, which may differ, due to numerous factors such as the reprogramming technology used. 

It is therefore important that we have simple, robust, and standardized techniques for evaluating and comparing individual iPSC lines, to ensure we choose the best lines for lengthy and expensive studies.

Conventional techniques for evaluating iPSC pluripotency and differentiation, such as traditional flow cytometry, have their drawbacks.

The development of alternative simple and robust methods, for monitoring iPSC differentiation has promise to enhance the scalability and throughput of applications utilizing iPSCs, such as liver toxicity screening.


Here we describe a combined iQue® Advanced Flow Cytometry and an Incucyte® Live-Cell Analysis approach for stem cell evaluation during cell line selection and differentiation. Hence creating a streamlined workflow for iPSC characterization, which measures differentiation marker expression over time. This method has potential applications in research and drug discovery.

  • Document type: Application Note
  • Page count: 9
  • Read time: 4 minutes

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Key Takeaways
  • iQue®Advanced Flow Cytometry Platform low volume requirements (10 μL)
  • Continuous morphological assessment, for easy monitoring of iPSC colonies throughout differentiation
  • Simple, one-wash labeling protocols 
  • Fast high-throughput sample acquisition
  • Plate-level, real-time data visualization tools for rapid comparison between cell lines
  • An easy way to check for pluripotency, confluency, and differentiation over time 

To ensure the best iPSC line was used for the differentiation protocol

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