High-density culture of vero cells in novel perfusion bioreactor system

Abstract

Improvements in upstream bioprocessing single-use systems have allowed implementation of high-density cultures in standard workflows. This study shows the capability of the novel designed single-use bioreactors to accommodates very high surface area per unit volume and enable the cell culture scientists to achieve and maintain a stable, high-density perfusion culture of Vero cells, without compromising mixing characteristics and nutritional homogeneity. Conventionally, adherent cell lines have been cultivated in non-controlled, labor-intensive, and space-demanding 2D surface technologies, i.e., roller bottles (RB) and multi-layer flasks. Fixed-bed and Packed-bed bioreactors are popular alternatives to culture high-density adherent cell lines for vaccines and other biologics production. However, the limitations that restricts the use of fixed-beds are non-homogeneous cell seeding, huge mass transfer gradients, inability of maintaining nutritional homogeneity, limitation of bed size expansion and difficulties during scale-up of etc. In this study, we have shown the design and development of novel disc-based bioreactors in single-use format for high-density Vero cell culture. During the process run of 12 days, we have observed 4.97 population doublings and final cell density achieved was 0.438 million cells/sq.cm from the seeding density of 0.014 million cells/sq.cm. Vero cell growth observed in novel bioreactor was significantly higher than the traditional culture devices, such as RB and multilayer flasks.