Large scale 3D bioreactor technology: linear scale up of mesenchymal stromal cell expansion with tidexcell™ system

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Background & Aim Tissue and bone marrow aspirate generally contain only a small amount of mesenchymal stromal cells (MSCs), yet a typical dose of 10-200 million cells or more is required for cell therapy. As such, alternatives to traditional planar culture platforms are preferred for efficient cell expansion. The extensive quantity demands for MSCs for allogeneic therapy further pushes the need for scaling up culture with bioreactors. In bioreactors, automation, single use and linearly scalable capabilities are desirable. However, this scaling up in bioreactors and bioprocesses is complex and requires extensive understanding experience and knowledge. Methods, Results & Conclusion Here, we performed upstream process development in a series of Tide Motion bioreactors that offer a culture environment with high oxygen transfer, low shear stress and no sparging. P4 bone marrow MSCs were recovered in planar cultures, seeded into a small benchtop bioreactor, the CelCradle, and harvested and reseeded into the TideXcell™ bioreactor with 1 L packed bed volume. Within a culture period of 15 days, the cells expanded by 9.6 doubling population levels, achieving 2 billion cells from 2.5 million cells. Real time monitoring of media/metabolites and sampling of cells ensured the culture was at optimal conditions throughout cell growth. Using the automated, closed TideXcell™ Cell Harvesting System, 97.7% harvesting efficiency was achieved with a cell viability of 93.8%. The criterial quality parameters (identity, purity, potency and safety) of the final cell product were inspected. As per ISCT recommendations, the MSCs demonstrated trilineage differentiation and immunophenotyping of > 95% for CD105, CD90, CD73, and <2% for CD34, CD45, HLA-DR, CD11b and CD19. T-cell suppression assay and IDO quantification also demonstrated intact immunomodulatory capabilities of the expanded MSCs. By coupling the automated TideXcell™ system together with its closed automated cell harvester, we were able to obtain high cell numbers with low labour cost. The bioprocess route to determining optimal culture parameters for the TideXcell™ was simplified by optimizing culture parameters in CelCradle™, with a 0.1 L packed-bed volume, before moving to larger capacities. The cell yield obtained in a 1 L packed bed TideXcell system was as projected from the CelCradle™ system. As such, the cost of bioprocess development was greatly reduced using the Tide Motion bioreactos. Tissue and bone marrow aspirate generally contain only a small amount of mesenchymal stromal cells (MSCs), yet a typical dose of 10-200 million cells or more is required for cell therapy. As such, alternatives to traditional planar culture platforms are preferred for efficient cell expansion. The extensive quantity demands for MSCs for allogeneic therapy further pushes the need for scaling up culture with bioreactors. In bioreactors, automation, single use and linearly scalable capabilities are desirable. However, this scaling up in bioreactors and bioprocesses is complex and requires extensive understanding experience and knowledge. Here, we performed upstream process development in a series of Tide Motion bioreactors that offer a culture environment with high oxygen transfer, low shear stress and no sparging. P4 bone marrow MSCs were recovered in planar cultures, seeded into a small benchtop bioreactor, the CelCradle, and harvested and reseeded into the TideXcell™ bioreactor with 1 L packed bed volume. Within a culture period of 15 days, the cells expanded by 9.6 doubling population levels, achieving 2 billion cells from 2.5 million cells. Real time monitoring of media/metabolites and sampling of cells ensured the culture was at optimal conditions throughout cell growth. Using the automated, closed TideXcell™ Cell Harvesting System, 97.7% harvesting efficiency was achieved with a cell viability of 93.8%. The criterial quality parameters (identity, purity, potency and safety) of the final cell product were inspected. As per ISCT recommendations, the MSCs demonstrated trilineage differentiation and immunophenotyping of > 95% for CD105, CD90, CD73, and <2% for CD34, CD45, HLA-DR, CD11b and CD19. T-cell suppression assay and IDO quantification also demonstrated intact immunomodulatory capabilities of the expanded MSCs.