Process Development for Expansion of Human Mesenchymal Stem Cells (hMSCs) using Three Dimensional (3D) Constructs in Novel Perfusion Bioreactors

摘要

Engineered tissue are at the cusp of a healthcare revolution in terms of their potential ability to regenerate damaged/lost tissue and organs. A key raw material for these products are the living cells, often required in the millions to billions of quantities for necessary function and replacement. This research focuses on the process development for the expansion of adherent cells, particularly adipose derived mesenchymal stem cells to meet the above demand. Two strategies were studied. In the first method, the therapeutic cells were encapsulated in 3D microgels produced through an electrostatic deposition process. In the second method, the adherent cells were attached to the surface of 3D printed polystyrene scaffolds inside novel perfusion bioreactors. Both these methods leverage the three-dimensional surface offered by the micro-environment, a significant advantage over standard 2D flat culture flasks in use today. Integration of the 3D printed porous scaffolds embedded within perfusion bioreactors were evaluated towards aiding the process of cell expansion and scale-up. This research has also contributed to the design and development of a novel tubeless perfusion bioreactor integrating 3D printed polystyrene scaffolds. The bioreactor was analyzed for critical process parameters and their optimization thereby laying the process development foundation for bioreactor design and operation for any adherent cell type. This research could lead to economically viable processes for adherent cell expansion leading to reduced cost in the manufacturability of Regenerative Medicine (RM) therapy products.