The application of MSC as cell therapeutics is a growing sector in health care. MSC therapeutics contain ~ 10^8 cells per dose, an amount that cannot solely be isolated from donor tissue. Thus, an in-vitro MSC expansion process is necessary and crucial, as it has a high impact on the quality of the final MSC product. The main goal of the project is the development of a novel dissolvable microcarrier that enables the adhesion and growth of mesenchymal stem cells (MSC) in a bioreactor process, and an efficient cell harvest while preserving the therapeutic potential of MSC. The porous, starch-based microcarriers will be optimized to ensure a mechanical support and a high surface-to-volume ratio when applied in a bioreactor. It will be dissolvable to enable a harvesting procedure that avoids a proteolytic detachment of the cells. A novel coating of the microcarriers will combine synthetic, bioactive peptides and glycosaminoglycans. The coating will be MSC-specific and will enable the adhesion and survival of the anchorage-dependent MSC in a serum-free cell culture medium, and support the preservation of their functionality. The microcarriers will be applied in a stirred-tank bioreactor. Process parameters promoting an optimal yield and MSC functionality will be identified and optimized in a design-of-experiments (DOE) approach. Physicochemical, cell-associated, and equipment-associated process parameters were investigated to determine critical ones. A limited fed batch model will be developed and then coupled with an online monitoring of the cell biomass via impedance spectroscopy. This process control strategy will be tested in various scenarios and for different MSC cell types. The novel microcarrier prototypes are tested in a real MSC expansion process and investigated regarding their sedimentation behaviour and their influence on mixing time and mass transfer processes, and analysed regarding the adhesion, growth, and the harvest of the MSC in the stirred tank reactor.
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