Monitoring of cell culture conditions and early prediction of the quality of an osteogenic cell-based medicinal product

Abstract

Background & Aim Novadip Biosciences developed a technology platform for tissue engineering and manufacturing of osteogenic cell-based pharmaceuticals. During the production process, a flexible medium refreshment strategy is essential to ensure the quality of the final product and the batch-to-batch consistency. The cellular metabolic activity of several dozens of batches was studied. A panel of metabolite concentrations was measured in cell culture supernatant from AMSCs isolation to formulation. Normalization of the data was carried out by calculating absolute and specific production/consumption rates. Methods, Results & Conclusion We defined optimal culture condition for AMSCs. In our setting, the level of metabolites stays in standard production/consumption rates and never reaches critically high values. A continuous consumption of glucose and glutamine is observed, and medium refreshment strategy allows to keep those nutrients at adequate level. A significant production of metabolites is observed throughout the process. The lactate level stays relatively low during the proliferation phase then raises after induction of osteogenic differentiation. It is showed that lactate that is a major end-product of glucose metabolism, reflects the transition between proliferating cells and osteogenic-oriented cells. Ammonia level tends to increase after osteo-induction but does not reach inhibitory concentration. Similarly, LDH level increases after osteo-induction. Glutamate consumption increases while pyruvate level decreases progressively during proliferation phase then go down to a low and stable level after osteoinduction. Phosphate concentration raises drastically during differentiation while the calcium level drops. Thus, the osteogenic differentiation leads to a metabolic shift in glucose, lactate, glutamate, LDH and pyruvate concentrations. Production and consumptions rates could be used to implement a routine-based analysis of waste media. Monitoring the rate of nutrients consumption and metabolites production represents a non-invasive and cost-effective strategy to monitor the process performance and predict quality issues with a high level of confidence.