Characterization and Optimization of Induced Pluripotent Stem Cell Culture Using Advanced Flow Cytometry and Live-Cell Analysis

Reading time: 13 minutes | Authors: Daryl Cole, Kirsty McBain, Nina Senutovitch, Nicola Bevan, Clare Szybut

Induced pluripotent stem cells (iPSCs) are a valuable resource in many areas of research and clinical development such as drug development, cancer research, organoid modeling of tissues and neurodevelopmental biology. In addition, iPSCs are increasingly used in translational applications. 

To maintain pluripotency, viability and propagation potential, iPSCs require highly specific optimal growth conditions. These requirements are often expensive and culture methods can be time intensive, requiring complicated techniques. 

In addition, long-term culture of iPSCs can result in genotypic and phenotypic heterogeneity, even in a cell line derived from a single source cell. Therefore, it is vital that methods for monitoring, detecting, and reducing heterogeneity in iPSC lines are developed.

This application note describes methods for the characterization, successful maintenance, scale up, and selection of ideal iPSCs in a combined workflow approach. This is achieved using rapid expression profiling with minimal sample volumes of 10 μL on the iQue® Advanced Flow Cytometry Platform, and real-time imaging on an Incucyte® Live-Cell Analysis System for easy morphological analysis.

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Key Takeaways

Key workflow advantages highlighted in this application note, include:

  • Streamlined processes and data analysis, which reduces attrition due to low-quality cell products
  • Miniaturization reducing antibody reagent costs and saving sample for downstream expansion, characterization, and differentiation
  • Monitoring iPSC lines throughout time courses, to provide greater insights into morphology and pluripotency changes over time
  • Rapid protocols requiring no fixation, in a single wash workflow for pluripotent surface markers

Figure 1. Schematic Highlighting the Combined iQue® and Incucyte® Workflow for iPSC Line Selection and Differentiation Monitoring

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