Stem cells have great potential to be used for clinical and drug discovery purposes. Large-scale “industrialized” production of stem cells will be necessary to enable their advancement for human therapeutics and to effectively deliver the quantities needed for drug discovery screening and lead optimization. However, achieving this level of production while meeting rigorous quality standards will depend on having the right tools and production systems for cell culture, scale-up, and characterization. Our bioreactor-based manufacturing platform will allow you to produce a consistent and reproducible supply of cells in an efficient and cost-effective way.
Growth Kinetics of human Mesenchymal Stem Cells in the Mobius® CellReady 3 L Single-use Bioreactor
The Mobius® CellReady 3 L single-use bioreactor can successfully be used to grow hMSCs on microcarriers. The information provided here shows how cells can be seeded directly into the bioreactor without prior need to attach the cells to microcarriers. After 2 weeks of culture, cells reach densities greater than 2×105 cells/mL while maintaining their identity as shown by the surface expression of CD105, CD90 and CD73 and the absences of CD14, CD34 and CD45. In addition the ability to differentiate to the adipogenic, chondrogenic, and osteogenic lineages is retained after culture. This system provides a cost-effective approach for the production of clinical grade hMSCs and allows for a close monitoring of the process.
Considerations for Scale-Up – Investigations into the Effects of Shear Forces on Human Mesenchymal Stem Cells
One of the areas of interest is the effect of shear on stem cells especially when transitioning from a flat (2D) tissue culture to a microcarrier culture. We have studied shear in a quantifiable model and used the markers to analyze hMSCs grown in microcarrier cultures both in spinner flask and the Mobius® CellReady 3L bioreactor with various agitation rates. This approach allows for a tight monitoring of shear forces in bioreactor cultures which is critical for a successful scale-up process.
Identity and Purity Characterization of MSCs Expanded in a 3L Stirred Tank Bioreactor
Mesenchymal stem cells (MSCs) are one of the key cell types being used for clinical study as therapeutic agents. Key parameters of product quality are identity and purity. A routine panel of assays typically used by industry was employed to characterize the cell state and the identity and purity of the cells. Cell state assays include viability, apoptosis and cell cycle. Identity and purity were assessed using both the standard recommended ISCT panel of positive and negative markers in addition to markers indicative of MSC properties. A combination of cell state and marker panels for identity and purity can be considered as a process monitoring tool for expansion of MSCs in addition to a quality control assessment following expansion.