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Strategies for the expansion of human induced pluripotent stem cells as aggregates in single-use Vertical-Wheel™ bioreactors
- Source :
- Journal of Biological Engineering, Vol 13, Iss 1, Pp 1-14 (2019), Journal of Biological Engineering
- Publication Year :
- 2019
- Publisher :
- BMC, 2019.
-
Abstract
- Background Since their inception, human induced pluripotent stem cells (hiPSCs) have held much promise for pharmacological applications and cell-based therapies. However, their potential can only be realised if large numbers of cells can be produced reproducibly on-demand. While bioreactors are ideal systems for this task, due to providing agitation and control of the culture parameters, the common impeller geometries were not designed for the expansion of mammalian cells, potentially leading to sub-optimal results. Results This work reports for the first time the usage of the novel Vertical-Wheel single-use bioreactors for the expansion of hiPSCs as floating aggregates. Cultures were performed in the PBS MINI 0.1 bioreactor with 60 mL of working volume. Two different culture media were tested, mTeSR1 and mTeSR3D, in a repeated batch or fed-batch mode, respectively, as well as dextran sulfate (DS) supplementation. mTeSR3D was shown to sustain hiPSC expansion, although with lower maximum cell density than mTeSR1. Dextran sulfate supplementation led to an increase in 97 and 106% in maximum cell number when using mTeSR1 or mTeSR3D, respectively. For supplemented media, mTeSR1 + DS allowed for a higher cell density to be obtained with one less day of culture. A maximum cell density of (2.3 ± 0.2) × 106 cells∙mL− 1 and a volumetric productivity of (4.6 ± 0.3) × 105 cells∙mL− 1∙d− 1 were obtained after 5 days with mTeSR1 + DS, resulting in aggregates with an average diameter of 346 ± 11 μm. The generated hiPSCs were analysed by flow cytometry and qRT-PCR and their differentiation potential was assayed, revealing the maintenance of their pluripotency after expansion. Conclusions The results here described present the Vertical-Wheel bioreactor as a promising technology for hiPSC bioprocessing. The specific characteristics of this bioreactor, namely in terms of the innovative agitation mechanism, can make it an important system in the development of hiPSC-derived products under current Good Manufacturing Practices.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Expansion
Aggregates
Environmental Engineering
Vertical-Wheel bioreactors
Biomedical Engineering
01 natural sciences
Flow cytometry
Single-use bioreactors
03 medical and health sciences
010608 biotechnology
Bioreactor
medicine
Human Induced Pluripotent Stem Cells
Bioprocess
Molecular Biology
lcsh:QH301-705.5
Dextran sulfate
Single use
Average diameter
medicine.diagnostic_test
Chemistry
Research
Fed-batch
Human induced pluripotent stem cells
Cell Biology
030104 developmental biology
lcsh:Biology (General)
Biophysics
Maximum Cell Density
Subjects
Details
- Language :
- English
- ISSN :
- 17541611
- Volume :
- 13
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Engineering
- Accession number :
- edsair.doi.dedup.....9bf44cf2764e1ea3ea74f6f8f0d894ec