A systematic approach combining process optimization and clinically-relevant quality attributes to manufacture high potency mesenchymal stromal cells for clinical and veterinary applications

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Problem Although advances in bioreactor technology allow for scalable manufacturing of mesenchymal stromal cells (MSCs), many of these methods are borrowed from the vaccine and antibody industry, and do not prioritize cell functionality. Commercial-scale expansion of MSCs contributed to sub-optimal performance in late-phase trials for multiple companies. Enhanced manufacturing methods that produce scaleable and fuctional MSCs are needed to capitalize on the substantial investment in MSCs. Solution My lab has filed a patent to manufacture scalable 3D MSCs with enhanced anti-inflammatory properties. Using a statistically-based approach, we will systematically optimize bioprocess parameters to manufacture sufficient quantities and quality of therapeutically-relevant MSCs with enhanced anti-inflammatory properties, without increasing costs. First, using iterative Design of Experiment (DoE) and high throughput read-outs, combinations of bioprocess parameters will be modeled and tested for their relative impact on MSC yield, anti-inflammatory factors and cost. Additional processing parameters that affect the metabolic profile of the MSC cultures will be investigated and queried to further optimize combination of bioprocess parameters. Human and canine MSCs will be manufactured in large scales, and tested in domestic dogs with inflammatory disease to validate this approach to optimize bioprocess parameters. O1 Statistical modeling to optimize process parameters that simultaneously improve MSC yield and anti-inflammatory attributes Herein, we use Definitive Screening Designs (DSD) to model the effects of various permutations of bioprocess parameters on MSC yield, potency and calculated cost. A response surface map allows hierarchical clustering of combination of bioprocess parameters that maximize cell yield and potency at low costs. O2 Assess metabolic parameters read-outs to further optimize MSC bioprocess parameters Optimizing metabolic parameters and changes in MSC size and cellular structures will be conducted in this objective using Raman spectroscopy. These parameters can subsequently be monitored on-line in MSC manufacturing vessels providing feedback to rapidly “correct” process parameters in situ.O3 Validate scalability and therapeutic potency of MSCs in veterinary applications Human and canine MSCs will be manufactured in 1L scales using the optimized combination of bioprocess parameters. Canine AT-MSCs will be tested in domestic dogs with joint diseases.Significance The process optimization will result in large scale manufacturing of therapeutically-potent MSCs for effective clinical and veterinary applications in inflammatory diseases. This program offers training opportunities in statistical modeling, bioprocess optimization, cell manufacturing, and veterinary application for HQPs.