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Herein, two translational issues related to the development of cell therapy discoveries into therapeutic products, are addressed. Firstly, analysis of whether the manipulations required to transfer cells from cell culture conditions to a target tissue affect cellular characteristics was performed. It was shown that processing primary murine mesenchymal stem cells (MSCs) into a concentrated cell suspension, drawing them up into a syringe and immediately ejecting them, caused a significant viability decrease. Leaving the cells within the syringe chamber at room temperature for prolonged time periods caused to a further decrease in viability. However, cells that were viable post-ejection were found to be functional with regard to their ability to attach and proliferate. Reducing the ejection rate or using the antioxidant n-acetyl cysteine did not significantly improve viability, although using a wider bore 22g needle did improve viability. Secondly, the feasibility of a cell transportation device that could store viable cells under room conditions was assessed. The development of such a transportation device would remove the need to cyropreserve cells during transit, which has many flaws. Human MSCs were found to enter a reversible proliferative arrest phase whilst under room conditions, allowing them to be stored for up to 11 days, before rapidly decreasing in viability. The accumulation of ammonia was identified within the cultures, and the introduction of a zeolite material was found to partially remove this ammonia and improve viability over 7 days. A means of developing a concentrated cell culture media, using a freeze-drying technique, was found not to comprise cell viability, whilst allowing the nutrient volume to be reduced, thus potentially making the transportation device more compact. However, the introduction of a modified release nutrient gel, consisting of alginate, did not provide any significant effect on viability. |
