Optimizing the production of suspension cells using the G-Rex “M” series
Author(s) -
Pradip Bajgain,
Roopa Mucharla,
John Wilson,
D.P. Welch,
Usanarat Anurathapan,
Bitao Liang,
Xiaohua Lu,
Kyle Ripple,
John M. Centanni,
Christine Hall,
David S. Hsu,
Larry A. Couture,
Shubhranshu S. Gupta,
Adrian P. Gee,
Helen E. Heslop,
Ann M. Leen,
Cliona M. Rooney,
Juan F. Vera
Publication year - 2014
Publication title -
molecular therapy — methods and clinical development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.285
H-Index - 32
ISSN - 2329-0501
DOI - 10.1038/mtm.2014.15
Subject(s) - scalability , suspension culture , cell culture , series (stratigraphy) , series and parallel circuits , computer science , seeding , biological system , chemistry , biochemical engineering , engineering , biology , electrical engineering , paleontology , database , voltage , genetics , aerospace engineering
Broader implementation of cell-based therapies has been hindered by the logistics associated with the expansion of clinically relevant cell numbers ex vivo. To overcome this limitation, Wilson Wolf Manufacturing developed the G-Rex, a cell culture flask with a gas-permeable membrane at the base that supports large media volumes without compromising gas exchange. Although this culture platform has recently gained traction with the scientific community due to its superior performance when compared with traditional culture systems, the limits of this technology have yet to be explored. In this study, we investigated multiple variables including optimal seeding density and media volume, as well as maximum cell output per unit of surface area. Additionally, we have identified a novel means of estimating culture growth kinetics. All of these parameters were subsequently integrated into a novel G-Rex “M” series, which can accommodate these optimal conditions. A multicenter study confirmed that this fully optimized cell culture system can reliably produce a 100-fold cell expansion in only 10 days using 1L of medium. The G-Rex M series is linearly scalable and adaptable as a closed system, allowing an easy translation of preclinical protocols into the good manufacturing practice
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