Scalable expansion of iPSC and their derivatives across multiple lineages
Chee Keong Kwok, Isabelle Sébastien, Krithika Hariharan, Ina Meiser, Jeanette Wihan, Saskia Altmaier, Isabell Karnatz, Dominic Bauer, Benjamin Fischer, Alexander Feile, Alfredo Cabrera‐Socorro, Mikkel A. Rasmussen, Bjørn Holst, Julia C. Neubauer, Christian Clausen, Catherine M. Verfaillie, Andreas Ebneth, Mattias Hansson, Rachel Steeg, Heiko Zimmermann
Abstract
Induced pluripotent stem cell (iPSC) technology enabled the production of pluripotent stem cell lines from somatic cells from a range of known genetic backgrounds. Their ability to differentiate and generate a wide variety of cell types has resulted in their use for various biomedical applications, including toxicity testing. Many of these iPSC lines are now registered in databases and stored in biobanks such as the European Bank for induced pluripotent Stem Cells (EBiSC), which can streamline the quality control and distribution of these individual lines. To generate the quantities of cells for banking and applications like high-throughput toxicity screening, scalable and robust methods need to be developed to enable the large-scale production of iPSCs. 3D suspension culture platforms are increasingly being used by stem cell researchers, owing to a higher cell output in a smaller footprint, as well as simpler scaling by increasing culture volume. Here we describe our strategies for successful scalable production of iPSCs using a benchtop bioreactor and incubator for 3D suspension cultures, while maintaining quality attributes expected of high-quality iPSC lines. Additionally, to meet the increasing demand for "ready-to-use" cell types, we report recent work to establish robust, scalable differentiation protocols to cardiac, neural, and hepatic fate to enable EBiSC to increase available research tools.