Process Development for the Expansion of Equine Cord Blood Mesenchymal Stem Cells in Stirred Suspension Bioreactors for the Treatment of Joint Injuries in Horses
Date
2018-04-30
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Abstract
Musculoskeletal injuries are the leading cause of lameness and loss of performance in horses with conventional treatments having high rates of re-injury. Equine Mesenchymal stem cells (eMSCs), specifically equine cord blood derived MSCs (eCB-MSCs) are a promising treatment alternative owing to their ability to differentiate into cartilage, as well as due to their immunomodulating properties. However, treatments can require cell numbers in the millions or billions, therefore conventional expansion methods using static T-flasks are typically inadequate in achieving these cell numbers. Expansion of eCB-MSCs in stirred suspension bioreactors with microcarriers as an attachment surface has the potential to obtain a clinically relevant number of cells while decreasing cost, time and labour requirements and increasing reproducibility and yield when compared to static expansion. As eCB-MSCs had not yet been expanded in stirred suspension bioreactors, a robust protocol was required to expand these cells using this method. This study outlines the process development of expansion, detailing the inoculation phase, the expansion phase, and the harvesting phase, followed by further testing on different donor cells. The process achieved maximum cell densities up to 75,000 cells/cm2 corresponding to 40 million cells in a 100mL bioreactor, with a harvesting efficiency of up to 80%, corresponding to a yield of 32 million cells in a 100mL bioreactor. Further testing is required to ensure eCB-MSCs maintained phenotype and functionality, however this study lays the groundwork for the large scale bioprocess of the expansion of eCB-MSCs.
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Stem Cells, Bioreactors, Regenerative Medicine
Citation
Roberts, E. L. (2018). Process Development for the Expansion of Equine Cord Blood Mesenchymal Stem Cells in Stirred Suspension Bioreactors for the Treatment of Joint Injuries in Horses (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/31900