Stem cell research is a rapidly growing field of study with huge potential for clinical solutions, taking advantage of the power and flexibility of pluripotency and self-renewal. Induced pluripotent stem cells (iPSCs) are capable of differentiating into all somatic cells of the body, whilst being infinitely expandable in culture. These characteristics make iPSCs not only uniquely valuable as a tool for developing culture models for the study of diseases and drug development, but also for the development of personalized therapeutics.
Generation of culture models and clinical therapies requires large quantities of healthy, pluripotent iPSCs with characteristics amenable to proliferation and subsequent differentiation. Maintenance of such cultures necessitates methodologies for the characterization and monitoring of key phenotypic markers, to ensure the preservation of pluripotency and homogeneity during growth. This becomes much more critical in the development of clinical applications, where safety is paramount for the patient. Having the ability to robustly monitor characteristics of iPSC cultures gives rise to opportunities for process design improvements during early development stages.
During the culturing of iPSCs, it is crucial to implement an environment that provides the precise concentrations and delivery of the various growth factors and cytokines which are vital for optimal expansion. Growth factors and cytokines in culture media will inevitably degrade over time, and at differing rates, so optimization of these key components is a requisite for maintenance of homogeneity during growth and expansion. The development of stable variants of growth factors such as FGF2, allow for reduced regularity of media changes, while maintaining stable concentrations of the molecule in culture media, helping to reduce the potential for heterogeneity. Further, various growth factors and cytokines are required at varying concentrations during iPSC differentiation, making the use of stable and high[1]quality supplements even more important when optimizing iPSC workflows.
This eBook provides a collection of data and methods for culturing, optimization, and characterization of iPSCs using flow cytometry as an analysis tool. From the optimization of iPSC culture and differentiation workflows to advanced methods for the characterization of surface and intracellular marker expression, the data presented here outline techniques and best practice for iPSC characterization using flow cytometry.
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