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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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Cites
Vascularized and functional human liver from an iPSC-derived organ bud transplant
Takebe, T., Sekine, K., Enomura, M., Koike, H., Kimura, M., Ogaeri, T., Zhang, R.-R., Ueno, Y., Zheng, Y.-W., Koike, N., et al. (2013) · Nature · doi:10.1038/nature12271 · Cited by 2,170
The promise of induced pluripotent stem cells in research and therapy
Robinton, D. A., & Daley, G. Q. (2012) · Nature · doi:10.1038/nature10761 · Cited by 1,150
Low O 2 tensions and the prevention of differentiation of hES cells
Ezashi, T., Das, P., & Roberts, R. M. (2005) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0501283102 · Cited by 824
Energy Metabolism in Human Pluripotent Stem Cells and Their Differentiated Counterparts
Varum, S., Rodrigues, A. S., Moura, M. B., Momcilovic, O., Easley, C. A., Ramalho-Santos, J., Van Houten, B., & Schatten, G. (2011) · PLoS ONE · doi:10.1371/journal.pone.0020914 · Cited by 671
UCP2 regulates energy metabolism and differentiation potential of human pluripotent stem cells
Zhang, J., Khvorostov, I., Hong, J. S., Oktay, Y., Vergnes, L., Nuebel, E., Wahjudi, P. N., Setoguchi, K., Wang, G., Do, A., et al. (2011) · The EMBO Journal · doi:10.1038/emboj.2011.401 · Cited by 512
Metabolic Regulation in Pluripotent Stem Cells during Reprogramming and Self-Renewal
Zhang, J., Nuebel, E., Daley, G. Q., Koehler, C. M., & Teitell, M. A. (2012) · Cell stem cell · doi:10.1016/j.stem.2012.10.005 · Cited by 477
Cho, Y. M., Kwon, S., Pak, Y. K., Seol, H. W., Choi, Y. M., Park, D. J., Park, K. S., & Lee, H. K. (2006) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2006.08.020 · Cited by 465
The promise of human induced pluripotent stem cells for research and therapy
Nishikawa, S., Goldstein, R. A., & Nierras, C. R. (2008) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm2466 · Cited by 423
Process engineering of human pluripotent stem cells for clinical application
Serra, M., Brito, C., Correia, C., & Alves, P. M. (2012) · Trends in biotechnology · doi:10.1016/j.tibtech.2012.03.003 · Cited by 309
Self-citationScalable expansion of human pluripotent stem cells in suspension culture
Zweigerdt, R., Olmer, R., Singh, H., Haverich, A., & Martin, U. (2011) · Nature Protocols · doi:10.1038/nprot.2011.318 · Cited by 271
Scalable Stirred-Suspension Bioreactor Culture of Human Pluripotent Stem Cells
Kehoe, D. E., Jing, D., Lock, L. T., & Tzanakakis, E. S. (2010) · Tissue Engineering Part A · doi:10.1089/ten.tea.2009.0454 · Cited by 264
Glutamine Oxidation Is Indispensable for Survival of Human Pluripotent Stem Cells
Tohyama, S., Fujita, J., Hishiki, T., Matsuura, T., Hattori, F., Ohno, R., Kanazawa, H., Seki, T., Nakajima, K., Kishino, Y., et al. (2016) · Cell Metabolism · doi:10.1016/j.cmet.2016.03.001 · Cited by 263
Saretzki, G. C., Walter, T., Atkinson, S. P., Passos, J. F., Bareth, B., Keith, W. N., Stewart, R., Hoare, S. F., Stojković, M., Armstrong, L., et al. (2007) · Stem Cells · doi:10.1634/stemcells.2007-0628 · Cited by 262
Human liver cell spheroids in extended perfusion bioreactor culture for repeated-dose drug testing
Tostões, R. M., Leite, S. B., Serra, M., Jensen, J., Björquist, P., Carrondo, M. J. T., Brito, C., & Alves, P. M. (2011) · Hepatology · doi:10.1002/hep.24760 · Cited by 255
Self-citationDerivation, propagation and controlled differentiation of human embryonic stem cells in suspension
Steiner, D., Khaner, H., Cohen, M. A., Even‐Ram, S., Gil, Y., Itsykson, P., Turetsky, T. T., Idelson, M., Aizenman, E., Ram, R., et al. (2010) · Nature Biotechnology · doi:10.1038/nbt.1616 · Cited by 232
Kempf, H., Olmer, R., Kropp, C., Rückert, M., Jara-Avaca, M., Diaz, D. R., Franke, A., Elliott, D. A., Wojciechowski, D., Fischer, M., et al. (2014) · Stem Cell Reports · doi:10.1016/j.stemcr.2014.09.017 · Cited by 215
Suspension Culture of Human Pluripotent Stem Cells in Controlled, Stirred Bioreactors
Olmer, R., Lange, A., Selzer, S., Kasper, C., Haverich, A., Martin, U., & Zweigerdt, R. (2012) · Tissue Engineering Part C Methods · doi:10.1089/ten.tec.2011.0717 · Cited by 207
Forsyth, N. R., Musio, A., Vezzoni, P., Simpson, H., Noble, B., & McWhir, J. (2006) · Cloning and Stem Cells · doi:10.1089/clo.2006.8.16 · Cited by 192
Serra, M., Correia, C., Malpique, R., Brito, C., Jensen, J., Björquist, P., Carrondo, M. J. T., & Alves, P. M. (2011) · PLoS ONE · doi:10.1371/journal.pone.0023212 · Cited by 179
Self-citationOlmer, R., Haase, A., Merkert, S., Cui, W., Paleček, J., Chen, R., Kirschning, A., Scheper, T. H., Glage, S., Miller, K., et al. (2010) · Stem Cell Research · doi:10.1016/j.scr.2010.03.005 · Cited by 172
Up-scaling single cell-inoculated suspension culture of human embryonic stem cells
Singh, H., Mok, P. Y., Balakrishnan, T., Rahmat, S. N. B., & Zweigerdt, R. (2010) · Stem Cell Research · doi:10.1016/j.scr.2010.03.001 · Cited by 157
Kropp, C., Kempf, H., Halloin, C., Diaz, D. R., Franke, A., Scheper, T. H., Kinast, K., Knorpp, T., Joos, T. O., Haverich, A., et al. (2016) · Stem Cells Translational Medicine · doi:10.5966/sctm.2015-0253 · Cited by 155
Serra, M., Brito, C., Sousa, M. F. Q., Jensen, J., Tostões, R. M., Clemente, J. J., Strehl, R., Hyllner, J., Carrondo, M. J. T., & Alves, P. M. (2010) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2010.06.015 · Cited by 150
Self-citationOtsuji, T. G., Jiang, B., Yoshimura, A., Tomura, M., Tateyama, D., Minami, I., Yoshikawa, Y., Aiba, K., Heuser, J. E., Nishino, T., et al. (2014) · Stem Cell Reports · doi:10.1016/j.stemcr.2014.03.012 · Cited by 136
Enterohepatic Helicobacter in Ulcerative Colitis: Potential Pathogenic Entities?
Thomson, J., Hansen, R., Berry, S. H., Hope, M. E., Murray, G. I., Mukhopadhya, I., McLean, M. H., Shen, Z., Fox, J. G., El-Omar, E. M., et al. (2011) · PLoS ONE · doi:10.1371/journal.pone.0017184 · Cited by 130
Ulvestad, M., Nordell, P., Asplund, A., Rehnström, M., Jacobsson, S., Holmgren, G., Davidson, L., Brolén, G., Edsbagge, J., Björquist, P., et al. (2013) · Biochemical Pharmacology · doi:10.1016/j.bcp.2013.06.029 · Cited by 108
Oxygen Transport and Stem Cell Aggregation in Stirred-Suspension Bioreactor Cultures
Wu, J., Rostami, M. R., Olaya, D. P. C., & Tzanakakis, E. S. (2014) · PLoS ONE · doi:10.1371/journal.pone.0102486 · Cited by 97
Chen, X., Chen, A., Woo, T. L., Choo, A. B. H., Reuveny, S., & Oh, S. K. W. (2010) · Stem Cells and Development · doi:10.1089/scd.2010.0077 · Cited by 89
Kerscher, P., Turnbull, I. C., Hodge, A. J., Kim, J., Seliktar, D., Easley, C. J., Costa, K. D., & Lipke, E. A. (2016) · Biomaterials · doi:10.1016/j.biomaterials.2015.12.011 · Cited by 88
Tostões, R. M., Leite, S. B., Miranda, J. P., Sousa, M. F. Q., Wang, D. I. C., Carrondo, M. J. T., & Alves, P. M. (2010) · Biotechnology and Bioengineering · doi:10.1002/bit.22920 · Cited by 86
Self-citationCorreia, C., Serra, M., Espinha, N., Sousa, M. F. Q., Brito, C., Burkert, K., Zheng, Y., Hescheler, J., Carrondo, M. J. T., Šarić, T., et al. (2014) · Stem Cell Reviews and Reports · doi:10.1007/s12015-014-9533-0 · Cited by 79
Self-citationDifferentiating Human Embryonic Stem Cells Express a Unique Housekeeping Gene Signature
Synnergren, J., Giesler, T. L., Adak, S., Tandon, R., Noaksson, K., Lindahl, A. H., Nilsson, P., Nelson, D. A., Olsson, B. E., Englund, M. C. O., et al. (2007) · Stem Cells · doi:10.1634/stemcells.2006-0247 · Cited by 63
Cunha, B., Aguiar, T., Silva, M. M., Silva, R. J. S., Sousa, M. F. Q., Pineda, E., Peixoto, C., Carrondo, M. J. T., Serra, M., & Alves, P. M. (2015) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2015.02.023 · Cited by 62
Self-citationSilva, M. M., Rodrigues, A. F., Correia, C., Sousa, M. F. Q., Brito, C., Coroadinha, A. S., Serra, M., & Alves, P. M. (2015) · Stem Cells Translational Medicine · doi:10.5966/sctm.2014-0270 · Cited by 40
Self-citationGuo, C., Kawakatsu, M., Idemitsu, M., Urata, Y., Goto, S., Ono, Y., Hamano, K., & Li, T. (2013) · Journal of Cellular Physiology · doi:10.1002/jcp.24389 · Cited by 35
Patient-specific hiPSC bioprocessing for drug screening: Bioprocess economics and optimisation
Jenkins, M., Bilsland, J., Allsopp, T. E., Ho, S. V., & Farid, S. S. (2015) · Biochemical Engineering Journal · doi:10.1016/j.bej.2015.09.024 · Cited by 28
Perfusion Stirred-Tank Bioreactors for 3D Differentiation of Human Neural Stem Cells
Simão, D. F., Arez, F., Terrasso, A. P., Pinto, C., Sousa, M. F. Q., Brito, C., & Alves, P. M. (2016) · Methods in molecular biology · doi:10.1007/7651_2016_333 · Cited by 26
Self-citationSart, S., Bejoy, J., & Li, Y. (2016) · Process Biochemistry · doi:10.1016/j.procbio.2016.05.024 · Cited by 24
Gomes‐Alves, P., Serra, M., Brito, C., Ricardo, C. P., Cunha, R. M. D. S., Sousa, M. F. Q., Sánchez, B., Bernad, A., Carrondo, M. J. T., Rodrı́guez-Borlado, L., et al. (2016) · Translational research · doi:10.1016/j.trsl.2016.02.001 · Cited by 16
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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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