Bioprocessing for Expansion and Standardization of Embryonic Stem Cell Cultures

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Since their derivation in 1998, human embryonic stem cells (ESCs) have been of great interest within the realm of regenerative medicine. Their ability to differentiate into any cell type of the adult body makes them an incredibly powerful and exciting tool with respect to potential treatments for long-term degenerative diseases. Stirred suspension bioreactors have been the focus of many publications anticipating the need for high numbers of clinically-relevant cells for therapeutic applications. As evidenced by the wide range of published protocols and resulting culture output, the field is currently lacking in well designed, systematic investigations into the true effect of varying culture parameters not only in suspension conditions but also within standard static cultures. This research was carried out to address gaps that currently exist within current ESC culture protocols first by establishing a base line for culture kinetics, during which the issue of morphological evaluation was identified. With the goal of eliminating the subjective nature of morphological evaluation, fractal geometry was utilized to assign a quantitative value (fractal dimension) to ESC cultures. Results indicate a significant difference between fractal dimensions determined for pluripotent cells versus differentiating cells suggesting that fractal geometry may be used to monitor culture health. A more standardized methodology for static culture monitoring will allow for more homogeneous population of ESCs as inoculum for large scale expansion efforts. Finally, a 3^2 factorial experimental design was developed and executed for stirred suspension culture of ESCs illustrating the potential for interaction effects and suggesting for the first time that the use of the conventional stepwise optimisation approach to stirred suspension culture is not appropriate.
Hunt, M. (2013). Bioprocessing for Expansion and Standardization of Embryonic Stem Cell Cultures (Doctoral thesis, University of Calgary, Calgary, Canada). Retrieved from doi:10.11575/PRISM/25139