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Effect of inoculum density on human-induced pluripotent stem cell expansion in 3D bioreactors
Charite Univ Med Berlin, Germany.
Charite Univ Med Berlin, Germany.
Charite Univ Med Berlin, Germany.
Univ Southern Calif, CA USA.
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2019 (English)In: Cell Proliferation, ISSN 0960-7722, E-ISSN 1365-2184, Vol. 52, no 4, article id e12604Article in journal (Refereed) Published
Abstract [en]

Objective For optimized expansion of human-induced pluripotent stem cells (hiPSCs) 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 x 10(6) or 50 x 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 (R) 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 x 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.

Place, publisher, year, edition, pages
WILEY , 2019. Vol. 52, no 4, article id e12604
Keywords [en]
3D culture; bioreactor culture; cell expansion; human-induced pluripotent stem cells; inoculum density
National Category
Cell Biology
Identifiers
URN: urn:nbn:se:liu:diva-159587DOI: 10.1111/cpr.12604ISI: 000477913100009PubMedID: 31069891OAI: oai:DiVA.org:liu-159587DiVA, id: diva2:1342294
Note

Funding Agencies|Bundesministerium fur Bildung und Forschung [13GW0129A]

Available from: 2019-08-13 Created: 2019-08-13 Last updated: 2019-08-13

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Mandenius, Carl-Fredrik
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