Current state and perspectives in modeling and control of human pluripotent stem cell expansion processes in stirred‐tank bioreactors

Galvanauskas, Vytautas; Grincas, Vykantas; Simutis, Rimvydas; Kagawa, Y.; Kino‐oka, Masahiro

doi:10.1002/btpr.2431

Cited by 10 publications

(5 citation statements)

References 80 publications

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“…Initially developed for use as an extracorporeal liver support system and successfully applied as such, 38 the compartmentalized bioreactor provides a countercurrent "arteriovenous" media flow and decentralized gas perfusion via capillaries, thereby enhancing mass exchange for an optimized nutrient and oxygen supply for the cultured cells. 39 Also, cells are not affected by shear stress, which occurs for example if cells are cultured in stirred tank bioreactors, 40 and can cause cell damage. 41 The total amount of cells produced in LS 50 (5.4 × 10 9 cells) would suffice for single-patient treatments in heart and liver therapies as well as treatment of diabetes.…”

Section: Discussionmentioning

confidence: 99%

Effect of inoculum density on human‐induced pluripotent stem cell expansion in 3D bioreactors

et al. 2019

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Objective: For optimized expansion of human-induced pluripotent stem cells (hiP-SCs) with regards to clinical applications, we investigated the influence of the inoculum density on the expansion procedure in 3D hollow-fibre bioreactors. Materials and Methods:Analytical-scale bioreactors with a cell compartment volume of 3 mL or a large-scale bioreactor with a cell compartment volume of 17 mL were used and inoculated with either 10 × 10 6 or 50 × 10 6 hiPSCs. Cells were cultured in bioreactors over 15 days; daily measurements of biochemical parameters were performed. At the end of the experiment, the CellTiter-Blue ® Assay was used for culture activity evaluation and cell quantification. Also, cell compartment sections were removed for gene expression and immunohistochemistry analysis. Results:The results revealed significantly higher values for cell metabolism, cell activity and cell yields when using the higher inoculation number, but also a more distinct differentiation. As large inoculation numbers require cost and time-extensive pre-expansion, low inoculation numbers may be used preferably for long-term expansion of hiPSCs. Expansion of hiPSCs in the large-scale bioreactor led to a successful production of 5.4 × 10 9 hiPSCs, thereby achieving sufficient cell amounts for clinical applications. Conclusions:In conclusion, the results show a significant effect of the inoculum density on cell expansion, differentiation and production of hiPSCs, emphasizing the importance of the inoculum density for downstream applications of hiPSCs. Furthermore, the bioreactor technology was successfully applied for controlled and scalable production of hiPSCs for clinical use.

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Section: Discussionmentioning

confidence: 99%

Effect of inoculum density on human‐induced pluripotent stem cell expansion in 3D bioreactors

et al. 2019

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“…Although pluripotent stem cells have been generated at laboratory scale, large scale production processes by standardized and economically viable procedures and technologies will be required 5 . One of the challenges in large scale expansion of pluripotent stem cells is suspension culture of cell aggregates in stirred bioreactors 6 , 7 in which single cells form cell aggregates in the presence of the small molecule Y27632 (Rho-associated coiled-coil kinase inhibitor) 8 . Furthermore, bioreactor-based suspension culture of pluripotent stem cells can be used for large scale induction of functional cells such as iPSC-derived cardiomyocytes 9 .…”

Section: Introductionmentioning

confidence: 99%

Magnetic heating of nanoparticles as a scalable cryopreservation technology for human induced pluripotent stem cells

Ito

Yoshioka

Masumoto

et al. 2020

Sci Rep

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Scale-up of production is needed for industrial applications and clinical translation of human induced pluripotent stem cells (hiPSCs). However, in cryopreservation of hiPSCs, successful rewarming of vitrified cells can only be achieved by convective warming of small volumes (generally 0.2 mL). Here, we present a scalable nano-warming technology for hiPSC cryopreservation employing inductive heating of magnetic nanoparticles under an alternating magnetic field. The conventional method by water bath heating at 37 °C resulted in a decrease of cell viability owing to devitrification caused by slow warming of samples with large volumes (≥ 20 mL). Nano-warming showed uniform and rapid rewarming of vitrified samples and improved viability of hiPSCs in the 20-mL system. In addition to single cells, hiPSC aggregates prepared using a bioreactor-based approach were successfully cryopreserved by the nano-warming technique. These results demonstrate that nano-warming is a promising methodology for cryopreservation in mass production of hipScs. Pluripotent stem cells, including human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiP-SCs), are a promising cell source for regenerative medicine because of their unlimited proliferation potential and differentiation capability 1,2. The first patient was treated with an hESC-based cellular therapy product in a clinical trial for spinal cord injury in 2010 3. In 2014, the first patient received hiPSC-derived retinal pigment epithelial cells for macular degeneration 4. Although pluripotent stem cells have been generated at laboratory scale, large scale production processes by standardized and economically viable procedures and technologies will be required 5. One of the challenges in large scale expansion of pluripotent stem cells is suspension culture of cell aggregates in stirred bioreactors 6,7 in which single cells form cell aggregates in the presence of the small molecule Y27632 (Rho-associated coiled-coil kinase inhibitor) 8. Furthermore, bioreactor-based suspension culture of pluripotent stem cells can be used for large scale induction of functional cells such as iPSC-derived cardiomyocytes 9. In addition, a recent report suggests that cell aggregates can be used as building blocks for tissue engineering 10. One bottleneck in manufacturing pluripotent stem cells is robust cryopreservation, and large-scale cell cryopreservation will be mandatory for industrial applications and clinical translation. Cryopreservation is classified into two distinct methods: slow freezing and vitrification. In vitrification methods, cryoprotectant solutions with high cryoprotectant concentrations are used, and cells are rapidly frozen by direct immersion of the container in liquid nitrogen. Originally, vitrification methods were developed for cryopreservation of oocytes and embryos 11. Fujioka et al. developed DAP213, a cryoprotectant solution containing dimethyl sulfoxide (DMSO) for primate ESCs, and succeeded in cryopreservation of ESCs by vitrification of a 0.2-mL scale 12. H...

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“…Depending on the therapeutic target, it is necessary to generate a quantity of 10 5 –10 10 cells [4]. Therefore a robust and high-performance culture system for successful cell expansion (maintaining efficient cell growth and preventing spontaneous differentiation) is essential, to provide sufficient quantities of undifferentiated human pluripotent stem cells (hPSCs) for further processing and differentiation [5]. Analysis of various cell cultivation technologies indicates that three-dimensional (3-D) suspension cultures in stirred-tank bioreactors provide good process scalability, controllability, and monitoring options [6], [7].…”

Section: Introductionmentioning

confidence: 99%

“…Galvanauskas et al. [5] have presented an extensive overview of existing modeling methods and techniques. To date, only a limited number of mathematical models [13], [14], [15] have been applied to optimization and control problems in stem cell expansion processes.…”

Section: Introductionmentioning

confidence: 99%

Kinetic modeling of human induced pluripotent stem cell expansion in suspension culture

Galvanauskas

Simutis

Nath

et al. 2019

Regenerative Therapy

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To date, practical application of mathematical models for model-based design of stem cell expansion processes is limited. Nevertheless, the first attempts show vast potential of this approach for the improvement of expansion process performance. This article presents the developed dynamic kinetic model of the human induced pluripotent stem cell expansion process in suspension culture. The model predicts cell growth, consumption of glucose and production of lactic acid, as well as the average aggregate size. The latter process variable is of particular importance for achieving high cell density. By adding botulinum hemagglutinin, an E-cadherin inhibitor and subsequent aggregate break-up, one can significantly increase performance of cell expansion process. After defining the appropriate optimization criteria and additional modification of the model, the latter can be further applied for model-based optimization of the final cell concentration by calculating optimal aggregate break-up and glucose/glutamine feeding strategies.

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Current state and perspectives in modeling and control of human pluripotent stem cell expansion processes in stirred‐tank bioreactors

Cited by 10 publications

References 80 publications

Effect of inoculum density on human‐induced pluripotent stem cell expansion in 3D bioreactors

Effect of inoculum density on human‐induced pluripotent stem cell expansion in 3D bioreactors

Magnetic heating of nanoparticles as a scalable cryopreservation technology for human induced pluripotent stem cells

Kinetic modeling of human induced pluripotent stem cell expansion in suspension culture

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