Methods and Process Optimization for Large-Scale CAR T Expansion Using the G-Rex Cell Culture Platform

  • Josh LudwigEmail author
  • Mark Hirschel
Part of the Methods in Molecular Biology book series (MIMB, volume 2086)


The G-Rex cell culture platform is based on a gas-permeable membrane technology that provides numerous advantages over other systems. Conventional bioreactor platform technologies developed for large scale mammalian cell expansion are typically constrained by the mechanics of delivering oxygen to an expanding cell population. These systems often utilize complex mechanisms to enhance oxygen delivery, such as stirring, rocking, or perfusion, which adds to expense and increases their overall risk of failure. On the other hand, G-Rex gas-permeable membrane-based bioreactors provide a more physiologic environment and avoid the risk and cost associated with more complex systems. The result is a more robust, interacting cell population established through unlimited oxygen and nutrients that are available on demand. By removing the need to actively deliver oxygen, these bioreactors can hold larger medium volumes (more nutrients) which allows the cells to reach a maximum density without complexity or need for media exchange. This platform approach is scaled to meet the needs of research through commercial production with a direct, linear correlation between small and large devices. In the G-Rex platform, examples of cell expansion (9–14 day duration) include; CAR-T cells, which have atypical harvest density of 20–30 × 106/cm2 (or 2–3 × 109 cells in a 100 cm2 device); NK cells, which have a typical harvest density of 20–30 × 106/cm2 (or 2–3 × 109 cells in a 100 cm2 device) and numerous other cell types that proliferate without the need for intervention or complex processes normally associated with large scale culture. Here we describe the methods and concepts used to optimize expansion of various cell types in the static G-Rex bioreactor platform.

Key words

G-Rex CAR T expansion Process development Large scale T cell culture Suspension culture In vitro cell culture Gas-permeable membrane Bioreactor 


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Wilson Wolf CorporationSt. PaulUSA

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