Abstract
This chapter describes a detailed protocol on human pluripotent stem cells (hPSCs) cultivation as matrix-free cell-only aggregates in defined and xeno-free culture medium in stirred tank bioreactors (STBRs). Starting with a frozen stock pre-expanded on conventional culture dishes (2D), the ultimate process is performed in 150 mL culture scale in stirred tank bioreactors (3D) and is designed to produce up to 500 million pluripotent hPSC within 7 days. The culture strategy includes perfusion-based cell feeding facilitating process control, automation, and higher cell yields. Ultimately, this detailed protocol describes an important step for generating a defined starting cell population for directed lineage differentiation and subsequently fueling human cell-based assays and regenerative medicine approaches.
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Acknowledgments
This work was supported by StemBANCC (support from the Innovative Medicines Initiative joint undertaking under grant 115439-2, whose resources are composed of financial contribution from the European Union [FP7/2007-2013] and EFPIA companies’ in-kind contribution). RZ received funding from the German Research Foundation (DFG; including the Cluster of Excellence REBIRTH EXC 62/2 and grants ZW64/4-1, KFO311, and ZW64/7-1), the German Ministry for Education and Science (BMBF; grants: 13 N14086, 01EK1601A, and 01EK1602A) and by the European Union (TECHNOBEAT; grant 66724).
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Manstein, F., Halloin, C., Zweigerdt, R. (2019). Human Pluripotent Stem Cell Expansion in Stirred Tank Bioreactors. In: Mandenius, CF., Ross, J. (eds) Cell-Based Assays Using iPSCs for Drug Development and Testing. Methods in Molecular Biology, vol 1994. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9477-9_7
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DOI: https://doi.org/10.1007/978-1-4939-9477-9_7
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Online ISBN: 978-1-4939-9477-9
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