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Expansion of 3D human induced pluripotent stem cell aggregates in bioreactors: Bioprocess intensification and scaling-up approaches
Abstract
Human induced pluripotent stem cells (hiPSC) are attractive tools for drug screening and disease modeling and promising candidates for cell therapy applications. However, to achieve the high numbers of cells required for these purposes, scalable and clinical-grade technologies must be established. In this study, we use environmentally controlled stirred-tank bioreactors operating in perfusion as a powerful tool for bioprocess intensification of hiPSC production. We demonstrate the importance of controlling the dissolved oxygen concentration at low levels (4%) and perfusion at 1.3day -1 dilution rate to improve hiPSC growth as aggregates in a xeno-free medium. This strategy allowed for increased cell specific growth rate, maximum volumetric concentrations (4.7×10 6 cell/mL) and expansion factors (approximately 19 in total cells), resulting in a 2.6-fold overall improvement in cell yields. Extensive cell characterization, including whole proteomic analysis, was performed to confirm that cells' pluripotent phenotype was maintained during culture. A scalable protocol for continuous expansion of hiPSC aggregates in bioreactors was implemented using mechanical dissociation for aggregate disruption and cell passaging. A total expansion factor of 1100 in viable cells was obtained in 11days of culture, while cells maintained their proliferation capacity, pluripotent phenotype and potential as well as genomic stability after 3 sequential passages in bioreactors.
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“The apparent under appreciation of reporting measured environmental parameters is exemplified by published bioreactor experiments that report only the target levels of environmental parameters (e.g., Karst et al., 2016; Abecasis et al., 2017).”
From Toward Best Practices for Controlling Mammalian Cell Culture Environments (2022) · cites this paper as Abecasis et al., 2017
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Metadata
- Authors
- Abecasis, B., Aguiar, T., Arnault, É., Costa, R., Gomes-Alves, P., Aspegren, A., Serra, M., & Alves, P. M.
- Affiliations
- Instituto de Biologia Experimental e Tecnológica (PT)
- Takara (Sweden) (SE)
- Corresponding author
- Aspegren, Anders
- Serra, Margarida
- Venue
- Journal of Biotechnology
- Publisher
- Elsevier BV
- Published
- 2017-03
- Volume
- 246
- Pages
- 81-93
- ISSN
- 0168-1656 (ISSN-L)
- OpenAlex
- W2580933284
- Cited by
- 100
- References
- 40
- Citations by year
- 2026: 7 · 2025: 8 · 2024: 14 · 2023: 10 · 2022: 6 · 2021: 14 · 2020: 15 · 2019: 15 · 2018: 8 · 2017: 3
- FWCI
- 4.58
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Fundação para a Ciência e Tecnologia (FCT)-funded projects CARDIOSTEM (MITP-TB/ECE/0013/2013)
- CardioRegen (HMSP-ICT/0039/2013)
- iNOVA4Health (UID/Multi/04462/2013)
- FCT (SFRH/BD/52475/2013)
- Vinnova Grant
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Physical Sciences › Chemical Engineering › Fluid Flow and Transfer Processes › Fluid Dynamics and Mixing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- human induced pluripotent stem cells, cell aggregates, stirred tank bioreactor, perfusion bioreactor, bioprocess intensification, low dissolved oxygen, cell expansion, mechanical dissociation, pluripotency maintenance, genome stability, cell therapy
- MeSH terms
- Cell Differentiation, Cells, Cultured, Humans, Oxygen, Phenotype, Cell Culture Techniques, Bioreactors, Proteomics, Genomic Instability, Cell Proliferation, Induced Pluripotent Stem Cells
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