Objectives, benefits and challenges of bioreactor systems for the clinical-scale expansion of T lymphocyte cells

Baudequin, Timothée; Nyland, Robin; Ye, Hua

doi:10.1016/j.biotechadv.2021.107735

Cited by 18 publications

(25 citation statements)

References 100 publications

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“…Currently, many commercial bioreactors such as G-REX® systems, a variety of rocking-motion bioreactors, CliniMACS® Prodigy, etc. are created to address the need for large-scale, clinical settings [ 34 ].…”

Section: Discussionmentioning

confidence: 99%

Optimizing interleukin-2 concentration, seeding density and bead-to-cell ratio of T-cell expansion for adoptive immunotherapy

et al. 2021

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Background The successful ex vivo expansion of T-cells in great numbers is the cornerstone of adoptive cell therapy. We aimed to achieve the most optimal T-cell expansion condition by comparing the expansion of T-cells at various seeding densities, IL-2 concentrations, and bead-to-cell ratios. we first expanded the peripheral blood mononuclear cells (PBMCs) of a healthy donor at a range of 20 to 500 IU/mL IL-2 concentrations, 125 × 103 to 1.5 × 106 cell/mL, and 1:10 to 10:1 B:C (Bead-to-cell) ratios and compared the results. We then expanded the PBMC of three healthy donors using the optimized conditions and examined the growth kinetics. On day 28, CD3, CD4, and CD8 expression of the cell populations were analyzed by flow cytometry. Results T-cells of the first donor showed greater expansion results in IL-2 concentrations higher than 50 IU/mL compared to 20 IU/mL (P = 0.02). A seeding density of 250 × 103 cell/mL was superior to higher or lower densities in expanding T-cells (P = 0.025). Also, we witnessed a direct correlation between the B:C ratio and T-cell expansion, in which, in 5:1 and 10:1 B:C ratios T-cell significantly expanded more than lower B:C ratios. The results of PBMC expansions of three healthy donors were similar in growth kinetics. In the optimized condition, 96–98% of the lymphocyte population expressed CD3. While the majority of these cells expressed CD8, the mean expression of CD4 in the donors was 19.3, 16.5, and 20.4%. Conclusions Our methodology demonstrates an optimized culture condition for the production of large quantities of polyclonal T-cells, which could be useful for future clinical and research studies.

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Section: Discussionmentioning

confidence: 99%

Optimizing interleukin-2 concentration, seeding density and bead-to-cell ratio of T-cell expansion for adoptive immunotherapy

et al. 2021

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“…An advantage of G-Rex is that several vessels can be inserted in a single standard incubator, potentially allowing parallel expansions of multiple products with just one instrument. In contrast, other platforms require specific and quite expensive bioreactors in addition to single-use culture bags or vessels, and the machinery generally needs to be multiplied to be able to perform more than one expansion run in parallel [8,9,11]. This of course increases the costs significantly, which may not be sustainable for small or academic laboratories.…”

Section: Discussionmentioning

confidence: 99%

“…In the last 10 to 20 years, different types of bioreactors have been developed to allow reproducible and large-scale expansion of adherent or nonadherent cells in closed systems for clinical use [8À11]. Bioreactors, such as the Xuri (previously Biowave), Quantum-Cell Expansion System or Miltenyi Prodigy, are partially or fully automated, and rely on mechanical rocking or stirring of culture vessels to guarantee adequate distribution of nutrients and gas exchange during cell expansion [8,9,11]. Bioreactors are expensive, however, and usually allow production of only one cell batch at a time, with expansion of each batch lasting 1 to 3 weeks in the case of T cells [9,11].…”

Section: Introductionmentioning

confidence: 99%

“…Bioreactors, such as the Xuri (previously Biowave), Quantum-Cell Expansion System or Miltenyi Prodigy, are partially or fully automated, and rely on mechanical rocking or stirring of culture vessels to guarantee adequate distribution of nutrients and gas exchange during cell expansion [8,9,11]. Bioreactors are expensive, however, and usually allow production of only one cell batch at a time, with expansion of each batch lasting 1 to 3 weeks in the case of T cells [9,11]. This implies that cell factories need to have several bioreactors to produce more cell products in parallel, which can be a handicap, especially when urgent, patient-dedicated ATMPs are being produced for gravely ill patients, as in our case [9].…”

Section: Introductionmentioning

confidence: 99%

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Optimization of therapeutic T cell expansion in G-Rex device and applicability to large-scale production for clinical use

et al. 2022

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“…One of the potential drawbacks of the CliniMACS Prodigy systems is that it is a single-component machine, where the culture vessel and centrifuge are the same. This leads to slower processing of samples as simultaneous processing of multiple batches is not possible with a single machine (Baudequin et al, 2021). In the report by Mock et al (2016), the final composition of the product manufactured by CliniMACS Prodigy is similar to the product from the WAVE bioreactor.…”

Section: Manufacturing Technologymentioning

confidence: 99%

Influence of Culture Conditions on Ex Vivo Expansion of T Lymphocytes and Their Function for Therapy: Current Insights and Open Questions

Sudarsanam

Buhmann

Henschler

2022

Front. Bioeng. Biotechnol.

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Ex vivo expansion of T lymphocytes is a central process in the generation of cellular therapies targeted at tumors and other disease-relevant structures, which currently cannot be reached by established pharmaceuticals. The influence of culture conditions on T cell functions is, however, incompletely understood. In clinical applications of ex vivo expanded T cells, so far, a relatively classical standard cell culture methodology has been established. The expanded cells have been characterized in both preclinical models and clinical studies mainly using a therapeutic endpoint, for example antitumor response and cytotoxic function against cellular targets, whereas the influence of manipulations of T cells ex vivo including transduction and culture expansion has been studied to a much lesser detail, or in many contexts remains unknown. This includes the circulation behavior of expanded T cells after intravenous application, their intracellular metabolism and signal transduction, and their cytoskeletal (re)organization or their adhesion, migration, and subsequent intra-tissue differentiation. This review aims to provide an overview of established T cell expansion methodologies and address unanswered questions relating in vivo interaction of ex vivo expanded T cells for cellular therapy.

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Objectives, benefits and challenges of bioreactor systems for the clinical-scale expansion of T lymphocyte cells

Cited by 18 publications

References 100 publications

Optimizing interleukin-2 concentration, seeding density and bead-to-cell ratio of T-cell expansion for adoptive immunotherapy

Optimizing interleukin-2 concentration, seeding density and bead-to-cell ratio of T-cell expansion for adoptive immunotherapy

Optimization of therapeutic T cell expansion in G-Rex device and applicability to large-scale production for clinical use

Influence of Culture Conditions on Ex Vivo Expansion of T Lymphocytes and Their Function for Therapy: Current Insights and Open Questions

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