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Article

Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System

RoosterBio, Inc., 5295 Westview Drive, Suite 275, Frederick, MD 21703, USA
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Bioengineering 2020, 7(3), 73; https://doi.org/10.3390/bioengineering7030073
Received: 2 June 2020 / Revised: 2 July 2020 / Accepted: 5 July 2020 / Published: 8 July 2020
(This article belongs to the Special Issue Stem Cell Bioprocessing and Manufacturing)
Human mesenchymal stem/stromal cells (hMSCs) have been investigated and proven to be a well-tolerated, safe therapy for a variety of indications, as shown by over 900 registered hMSC-based clinical trials. To meet the commercial demand for clinical manufacturing of hMSCs, production requires a scale that can achieve a lot size of ~100B cells, which requires innovative manufacturing technologies such as 3D bioreactors. A robust suspension bioreactor process that can be scaled-up to the relevant scale is therefore crucial. In this study, we developed a fed-batch, microcarrier-based bioreactor process, which enhances media productivity and drives a cost-effective and less labor-intensive hMSC expansion process. We determined parameter settings for various stages of the culture: inoculation, bioreactor culture, and harvest. Addition of a bioreactor feed, using a fed-batch approach, was necessary to replenish the mitogenic factors that were depleted from the media within the first 3 days of culture. Our study resulted in an optimized hMSC culture protocol that consistently achieved hMSC densities between 2 × 105–6 × 105 cells/mL within 5 days with no media exchange, maintaining the final cell population doubling level (PDL) at 16–20. Using multiple hMSC donors, we showed that this process was robust and yielded hMSCs that maintained expansion, phenotypic characteristic, and functional properties. The developed process in a vertical-wheel suspension bioreactor can be scaled to the levels needed to meet commercial demand of hMSCs. View Full-Text
Keywords: bioreactor; hMSCs; microcarrier; bioprocess bioreactor; hMSCs; microcarrier; bioprocess
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MDPI and ACS Style

Lembong, J.; Kirian, R.; Takacs, J.D.; Olsen, T.R.; Lock, L.T.; Rowley, J.A.; Ahsan, T. Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System. Bioengineering 2020, 7, 73. https://doi.org/10.3390/bioengineering7030073

AMA Style

Lembong J, Kirian R, Takacs JD, Olsen TR, Lock LT, Rowley JA, Ahsan T. Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System. Bioengineering. 2020; 7(3):73. https://doi.org/10.3390/bioengineering7030073

Chicago/Turabian Style

Lembong, Josephine; Kirian, Robert; Takacs, Joseph D.; Olsen, Timothy R.; Lock, Lye T.; Rowley, Jon A.; Ahsan, Tabassum. 2020. "Bioreactor Parameters for Microcarrier-Based Human MSC Expansion under Xeno-Free Conditions in a Vertical-Wheel System" Bioengineering 7, no. 3: 73. https://doi.org/10.3390/bioengineering7030073

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