Culture and characterization of mesenchymal stem cells in a totally defined medium

Abstract

A successful transition from laboratory scale, to an efficient and robust good manufacturing practice (GMP) process is key and represents a major challenge for cell based products manufacturing. Implementing innovative technologies is essential to support such industrialization. Single-use bioreactors represent a solution by minimizing manual handling, realizing economies of scale and delivering the benefits of a closed system. However the implementation of bioreactor systems for large scale cell therapy industrialization is challenging. Scale-up is not as simple as providing a larger surface area. Changing the niche environment has a big impact on cell behaviour as cells are highly sensitive to the microenvironment. Also as the production scale increases, the volumes to be processed become substantial and require an adaptation of the downstream strategy. The combination of the Xpansion multiplate bioreactors and a continuous centrifuge enables an easy transfer from existing multitray stack process to a large scale and closed production system. The bioreactor offers the same cell growth environment (polystyrene multiplate) plus the regulation of the critical cell culture parameters as pH, dissolved oxygen and cell density. Single-use centrifuge device enable continuous concentration, wash and resuspend cells after culture. The system respects constraints from the cell therapy application: low-shear stress necessary to process stem cells, capture very low cell suspension concentration and high concentration factor. This presentation illustrates with different cases studies the transition from traditional open R&D systems to a closed manufacturing platform. Trends analysis of pH and DO for several cell cultures, as well as cell confluence monitoring, highlight efficiency of controls and monitoring to develop a robust scalable process. Cell quality was assayed and showed equivalence between all scales. Value and benefits are also examined through COGs analysis.