Abstract
Cell expansion is typically a long and labor-intensive step in CAR-T cell manufacture. The Xuri Cell Expansion System (CES) W25 semiautomates this step while functionally closing the process. Cells for autologous or allogeneic cell therapies are cultured inside a single-use Xuri Cellbag™ bioreactor. Wave-induced agitation, performed by a rocking Base Unit, transfers gas and mixes the culture. The integral UNICORN™ software allows customization of culture conditions and media perfusion schedules. Culture volumes can range from 300 mL to 25 L, making the Xuri CES W25 system suitable for both scale-up and scale-out manufacturing processes. CAR-T cell therapies have been successfully generated using the Xuri CES W25 system, which reduces manual labor compared with static culturing methods. This chapter details how to initiate a culture, install the Xuri CES W25, and install a 2 L Cellbag bioreactor. Protocols on inoculation, monitoring, and sampling are also outlined in this chapter.
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References
Vormittag P, Gunn R, Ghorashian S, Veraitch F (2018) A guide to manufacturing CAR T cell therapies. Curr Opin Biotechnol 53:164–181
Janas ML, Nunes C, Marenghi A, Sauvage V, Davis B, Bajas A, Burns A (2015) Perfusion’s role in maintenance of high-density T-cell cultures. Bioprocess Int 13(1):18–26
Areman EM, Loper K (2009) Cellular therapy: principles, methods, and regulations. AABB Press, Bethesda
Hollyman D, Stefanski J, Przbylowski M, Bartido S, Borquez-Ojeda O, Taylor C et al (2009) Manufacturing validation of biologically functional T cells targeted to CD19. J Immunother 32(2):169–180
Sadeghi A, Pauler L, Anneré C, Friberg A, Brandhorst D, Korsgren O, Totterman TH (2011) Large-scale bioreactor expansion of tumor-infiltrating lymphocytes. J Immunol Methods 364:94–100
Healthcare GE (2018) A semi-automated, high-purity process for NK cell manufacturing in a rocking bioreactor. (KA4771180618PO). General Electric, Boston, MA
Davis BM, Loghin ER, Conway KR, Zhang X (2018) Automated closed-system expansion of pluripotent stem cell aggregates in a rocking-motion bioreactor. SLAS Technol 23(4):364–373. https://doi.org/10.1177/2472630318760745
Kokaji AI, Sun CA, Ng V, Lam BS, Clark SJ, Woodside SM, Eaves AC, Thomas TE (2016) Scalable human T cell isolation, activation and expansion using EasySep™ and ImmunoCult™ by STEMCELL Technologies. Eur J Immunol 46(Suppl 1):119–120
Brentjens RJ et al (2011) Safety and persistence of adoptively transferred autologous CD19-targeted T cells in patients with relapsed or chemotherapy refractory B-cell leukemias. Blood 118(18):4817–4828
Iyer RK, Bowles PA, Howard K, Dulgar-Tulloch A (2018) Industrializing autologous adoptive immunotherapies: manufacturing advances and challenges. Front Med 5(150):1–9
GE Healthcare (2016) T cell expansion with Xuri™ systems: isolation and cultivation protocol (document number 29112375 revision AB). General Electric, Boston, MA
Golubovskaya V, Wu L (2016) Different subsets of T cells, memory, effector functions, and CAR-T immunotherapy. Cancers (Basel) 8(3):36
Xu W, Larbi A (2017) Markers of T cell senescence in humans. Int J Mol Sci 18(8):1742
Ismail R, Janas M, Stone S, Marenghi A, Sauvage V (2013) Expansion of T-cells using the Xuri™ cell expansion W25 and WAVE bioreactor™ 2/10 system. In: Developments in cell expansion for cell processing. GE Healthcare Life Sciences, Chicago, Illinois. https://www.gelifesciences.com/solutions/cell-therapy/knowledge-center/resources/developments-in-cell-expansion. Accessed 22 May 2018
Gee AP (2018) GMP CAR-T cell production. Best Pract Res Clin Haematol 31(2):126–134
Acknowledgments
The author thanks Matt Sherman, PE for providing the 2 L Cellbag bioreactor line drawing in Fig. 2, the Cellular Biology team at the GE Global Research Center for their continued mentorship, and the Centre for Commercialization of Regenerative Medicine for their persistence in expanding the applications of the Xuri Cell Expansion System W25.
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Smith, T.A. (2020). CAR-T Cell Expansion in a Xuri Cell Expansion System W25. In: Swiech, K., Malmegrim, K., Picanço-Castro, V. (eds) Chimeric Antigen Receptor T Cells. Methods in Molecular Biology, vol 2086. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0146-4_11
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DOI: https://doi.org/10.1007/978-1-0716-0146-4_11
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0146-4
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