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David Kuninger

Thermo Fisher Scientific

Director, R&D and Lead of Cell Models within the Cell Biology Business of Thermo Fisher Scientific.

Seasoned Biotechnology professional with broad experience driving innovation, product development and commercialization. Currently lead multiple R&D teams supporting Stem Cell Culture and Differentiation, Neurobiology, Primary Cells & ADME/Tox, 3D Biology and Cell Therapy applications incorporating these cell types. In the last 5 years his group has been responsible for the launch of over 40 new culture system products spanning a diverse array of biological applications.

 Differentiation of iPSCs in 3D: Leveraging Suspension Cultures for Scale and Efficiency

Abstract:

As the use of pluripotent stem cells (PSCs) in therapeutic and screening applications continues to expand, a key bottleneck is in the efficient generation high-quality PSCs. Three-dimensional (3D) suspension cultures offer several key advantages for scale up over two-dimensional (2D) adherent culture including overall cost, reduced footprint and hands on time, as well as compatibility with closed systems and reduced risk of contamination.   In addition, expansion of PSCs in 3D cultures consumes less media (and plasticware) than the same number of cells grown in 2D cultures. Together, these features make 3D culture an attractive path for cost-effectively and rapidly generating large quantities of cells required for certain downstream applications. We have recently developed a new 3D suspension culture medium – Gibco™ StemScale Medium, which was designed to promote the efficient self-aggregation of singularized PSCS into spheroids, without the need for microcarriers, while maintaining robust growth per culture level.  Both viability and pluripotency remain high for spheroids cultured in this medium over consecutive passages.   Here we show these expanded iPSC spheroids can be taken directly into various directed differentiation protocols while maintained in 3D suspension culture.  Our results demonstrate highly efficient induction into all three germ lines and subsequent downstream generation of cardiomyocytes and neurons as specific examples.  Strategies and approaches for adapting protocols designed for adherent culture for use in 3D suspension applications as well as potential benefits of “toggling”  between 2D and 3D culture are discussed.