Optimization of culture conditions for human bone marrow‐derived mesenchymal stromal cell expansion in macrocarrier‐based Tide Motion system

Background With high cell doses required for mesenchymal stromal cell (MSC) clinical trials, there is a need to upgrade technologies that facilitate efficient scale up of MSCs for cell therapy. Conventional expansion with 2D culture vessels becomes the bottleneck when large cell dosages are required. Tide Motion bioreactors offer a robust, scalable platform using BioNOC II macrocarriers developed for the production of adherent cells. Methods We evaluated the growth and expansion of bone marrow‐derived MSCs (BM‐MSCs) on the macrocarrier‐based culture system by optimizing key parameters such as cell seeding densities, culturing conditions, and harvesting procedures to achieve optimal cell growth. BM‐MSCs expanded in conventional 2D adherent cultures were seeded into BioNOC II macrocarriers and grown in serum‐containing or serum‐free medium. Results BM‐MSCs attained a maximum cell density of 0.49 ± 0.07 × 106 cells/carrier after 12 days of culture in BioNOC II macrocarriers with cell viability > 86% while retaining MSC specific characteristics such as surface marker expression, tri‐lineage differentiation potential, immunosuppressive properties, and potency. Conclusion These results reveal the feasibility of BM‐MSC expansion in the scalable macrocarrier‐based Tide Motion system both under serum and serum‐free conditions and represent an important step for the large‐scale production system of BM‐MSC based cellular therapies. Successful scaling up of mammalian cell cultures in a commercial setting requires the potential optimization of various parameters. Here the impact of seeding density and media type on the expansion of mesenchymal stromal cells (MSCs) cultured in BioNOC II macrocarriers is analyzed. Successful expansion of MSCs from a low seeding density in both serum and serum‐free media is demonstrated, indicating the feasibility of utilizing BioNOC II microcarrier‐based tide motion system as a platform for high capacity manufacturing of MSCs.