Growth of Mesenchymal Stromal Cells in Automated Microwell Cultures: Influence of the Engineering Environment on Cell Growth Kinetics and Non-Directed Differentiation

Human Mesenchymal Stromal Cells (hMSC) have the potential to differentiate into lineages of mesoderm origin, such as osteogenic, chondrogenic, and adipogenic lineages, presenting a promising potential for autologous regenerative medicine applications. However, one of the major challenges associated with delivering hMSC to the clinic is the propagation of undifferentiated cells in vitro in order to reach the quantity required for therapeutic applications. This thesis investigates the effect of environmental parameters that impact cell growth characteristics of hMSC in microwell plates, for the design of an automated cell expansion process in a robotic platform. As a result of this work, the main parameters that can be used to control the growth rate and differentiation potential of hMSC at all stages of the cell expansion process have been identified. A mathematical model has also been developed to forward predict the cell growth characteristic of hMSC for an individual donor, allowing for a patient specific bioprocess design that will ensure enough cells can be supplied back to the patient in a timely manner while assuring the quality of the final product. Process parameters for the cell expansion process of hMSC in automated microwells have been characterised. Optimum values for inoculation cell density and medium exchange strategies have been proven to reduce the overall time of the cell expansion process for hMSC in an undifferentiated state for their use in regenerative medicine therapies. These parameters were implemented in the cell expansion in an automated platform for the duration of one passage, proving the potential of such technology for the delivery of hMSC to the clinic.