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Abstract <p>Efficient large-scale expansion of adherent mammalian cells is essential for the production of recombinant proteins, viral vectors, and vaccines. Conventional multilayer flasks rely on scale-out approaches, require extensive manual handling, and lack monitoring process control compared to bioreactors. Here, we evaluate the CellScrew®, a dynamic 2D culture system available in three sizes ranging from 851 to 10,197 cm<sup>2</sup>, in which cell culture surfaces are continuously rotated, generating controlled hydrodynamic conditions that enhance mass transfer while reducing material consumption. Hydrodynamic conditions and oxygen transfer were characterized, with k<sub>L</sub>a values up to 6.69 ± 0.10&#xa0;h<sup>−1</sup>. In this context, we assessed cell attachment, proliferation, metabolism, and harvest performance using human embryonic kidney 293 (HEK-293), murine melanoma (B16-F10), and murine colon carcinoma (CT-26 WT) cells, which serve as established model systems for biotherapeutic manufacturing applications such as viral vectors and vaccine production. The system supported cell densities up to 7.7 × 10E5 cells per cm<sup>2</sup> (HEK-293), 8.7 × 10E4 cells per cm<sup>2</sup> (B16-F10), and 1.1 × 10E5 cells per cm<sup>2</sup> (CT-26 WT) with viabilities above 92%, comparable to multilayer flasks. Glucose consumption and lactate accumulation indicated stable metabolic activity. Mechanical harvesting enabled recovery efficiencies exceeding 89% while reducing detachment reagent usage by 67%. Overall, these results suggest that the CellScrew® can support robust adherent cell expansion with potential improvements in process efficiency and reduced resource consumption, indicating that it may represent a scalable and potentially more sustainable alternative to traditional multilayer platforms.</p> Key points <p>• <i>CellScrew® reaches up to 0.77 ± 0.14 × 10E6 cells per cm</i><sup><i>2</i></sup><i> (cell-line dependent).</i></p> <p>• <i>Harvest efficiency with 67% less detachment reagent use.</i></p> <p>• <i>High oxygen transfer (kLa up to 6.69 ± 0.10 h−1) enables efficient culture.</i></p>

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Characterization and evaluation of a novel polylactic acid-made bioreactor for large-scale adherent cell expansion

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月22日

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  • Björn Boshof,
  • Alena Hüppner,
  • Johanna Eichberg,
  • Henning Reyer,
  • Lukas Käßer,
  • Anne Kölsch

摘要

Abstract

Efficient large-scale expansion of adherent mammalian cells is essential for the production of recombinant proteins, viral vectors, and vaccines. Conventional multilayer flasks rely on scale-out approaches, require extensive manual handling, and lack monitoring process control compared to bioreactors. Here, we evaluate the CellScrew®, a dynamic 2D culture system available in three sizes ranging from 851 to 10,197 cm2, in which cell culture surfaces are continuously rotated, generating controlled hydrodynamic conditions that enhance mass transfer while reducing material consumption. Hydrodynamic conditions and oxygen transfer were characterized, with kLa values up to 6.69 ± 0.10 h−1. In this context, we assessed cell attachment, proliferation, metabolism, and harvest performance using human embryonic kidney 293 (HEK-293), murine melanoma (B16-F10), and murine colon carcinoma (CT-26 WT) cells, which serve as established model systems for biotherapeutic manufacturing applications such as viral vectors and vaccine production. The system supported cell densities up to 7.7 × 10E5 cells per cm2 (HEK-293), 8.7 × 10E4 cells per cm2 (B16-F10), and 1.1 × 10E5 cells per cm2 (CT-26 WT) with viabilities above 92%, comparable to multilayer flasks. Glucose consumption and lactate accumulation indicated stable metabolic activity. Mechanical harvesting enabled recovery efficiencies exceeding 89% while reducing detachment reagent usage by 67%. Overall, these results suggest that the CellScrew® can support robust adherent cell expansion with potential improvements in process efficiency and reduced resource consumption, indicating that it may represent a scalable and potentially more sustainable alternative to traditional multilayer platforms.

Key points

• CellScrew® reaches up to 0.77 ± 0.14 × 10E6 cells per cm2 (cell-line dependent).

• Harvest efficiency with 67% less detachment reagent use.

• High oxygen transfer (kLa up to 6.69 ± 0.10 h−1) enables efficient culture.