Strategies for the expansion of human induced pluripotent stem cells as aggregates in single-use Vertical-Wheel™ bioreactors.
Diogo E S Nogueira,Carlos A V Rodrigues,Marta Carvalho,Cláudia C. Miranda,Yas Hashimura,Sunghoon Jung,Brian Lee,Joaquim M. S. Cabral +7 more
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TLDR
The results here described present the Vertical-Wheel bioreactor as a promising technology for hiPSC bioprocessing, namely in terms of the innovative agitation mechanism, which can make it an important system in the development of hiPSCs-derived products under current Good Manufacturing Practices.Abstract:
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. 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. 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.read more
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Scalable Production of Human Mesenchymal Stromal Cell-Derived Extracellular Vesicles Under Serum-/Xeno-Free Conditions in a Microcarrier-Based Bioreactor Culture System
Miguel de Almeida Fuzeta,Miguel de Almeida Fuzeta,Nuno Bernardes,Filipa D. Oliveira,Ana Costa,Ana Fernandes-Platzgummer,José Paulo S. Farinha,Carlos A V Rodrigues,Sunghoon Jung,Rong-Jeng Tseng,William Milligan,Brian Lee,Miguel A. R. B. Castanho,Diana Gaspar,Joaquim M. S. Cabral,Cláudia Lobato da Silva +15 more
TL;DR: It is demonstrated that this scalable culture system can robustly manufacture EVs from MSC derived from different tissue sources, toward the development of novel therapeutic products.
Journal ArticleDOI
Optimized serial expansion of human induced pluripotent stem cells using low-density inoculation to generate clinically relevant quantities in vertical-wheel bioreactors.
Breanna S. Borys,Tania So,James Colter,Tiffany Dang,Erin L. Roberts,Tamas Revay,Leila Larijani,Roman Krawetz,Ian A. Lewis,Bob Argiropoulos,Derrick E. Rancourt,Sunghoon Jung,Yas Hashimura,Brian Lee,Michael S. Kallos +14 more
TL;DR: It is demonstrated that a vertical‐wheel bioreactor‐based bioprocess can provide optimal conditions for efficient, rapid generation of high‐quality hiPSCs to meet the demands for clinical manufacturing of therapeutic cell products.
Journal ArticleDOI
Bioprocess Technologies that Preserve the Quality of iPSCs.
TL;DR: This review presents an overview of cell culture media, suspension modalities, and monitoring techniques that preserve the quality and pluripotency of iPSCs during initiation, expansion, and manufacturing.
Journal ArticleDOI
Transcriptome profiling of human pluripotent stem cell‐derived cerebellar organoids reveals faster commitment under dynamic conditions
Teresa Silva,Teresa Silva,Rui Sousa-Luís,Tiago G. Fernandes,Evguenia Bekman,Evguenia Bekman,Carlos A V Rodrigues,Sandra H. Vaz,Sandra H. Vaz,Leonilde M. Moreira,Yas Hashimura,Sunghoon Jung,Brian Lee,Maria Carmo-Fonseca,Joaquim M. S. Cabral +14 more
TL;DR: In this article, a reproducible and scalable production of cerebellar organoids by using the single-use Vertical-Wheel bioreactors was presented, in which functional cerebella neurons were obtained.
Journal ArticleDOI
Perspectives on scaling production of adipose tissue for food applications
TL;DR: In this article , the authors discuss the topic of growing adipocytes in vitro that could imbue meat alternatives with the complex flavor and aromas of animal meat, and outline potential paths for the large scale production of in vitro cultured fat, including adipogenic precursors during cell proliferation.
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