Introduction: Mesenchymal stromal cells (MSC) are emerging as a leading cellular therapy for a number of diseases. Whilst promising as a cellular therapy, in order for MSC based therapy to become economically viable, this manufacturing process will have to be automated in a closed system bioreactor.
Method: A packed bed bioreactor was fabricated using Polydimethylsiloxane (PDMS) for the walls of the bioreactor vessel and air plasma treated polystyrene scaffold was used to provide large continuous surface area for cell growth. To optimise the design and expansion methods Green fluorescent protein (GFP) labelled mouse MSC were used. IVIS imaging was performed using propidium iodide to stain the fixed cells attached scaffold to determine the seeding density throughout the bioreactor. Cell growth was determined by metabolic base assay using Almar blue.
Results: IVIS images show that an even distribution of cells was achieved throughout the bioreactor using seeding method devised. Approximately 3 population doublings (2,000 cells/cm2 to 15,000 cells/cm2) was achieved growing GFP labelled mouse MSC on the polystyrene scaffold in static and single pass perfusion conditions over 6 days of culture.
Conclusion: The packed bed bioreactor is a viable option for the provision of the large surface area required to generate clinical doses of MSC.