Barrios Silva, Lady Vanessa; (2025) Developing Cells-in-Gel systems: Implications of biomaterials in long-term cell storage, desiccation and cell therapy-based applications. Doctoral thesis (Ph.D), UCL (University College London).

Text Lady Barrios Silva PhD Thesis.pdf - Accepted Version Access restricted to UCL open access staff until 1 July 2026. Download (69MB)

Abstract

With the increasing demand for mammalian cells in clinical cell-based therapies, it is crucial to optimise delivery and storage practices. Cryopreservation is the standard approach for long-term storage; however, it has significant logistical, financial, biological and accessibility drawbacks. Cell desiccation offers an alternative solution, removing cold-chain supply dependency, reducing costs and making cell-based therapies more accessible to developing countries. This project sought to improve understanding of mammalian cell desiccation requirements and develop a viable protocol. Methods: Biomaterial were fully characterised via SEM, DSC, TGA, FTIR, Franz cells, drying profiles, biocompatibility, rheology and angiogenic effect via the CAM assay. Following this, systematic experimentation (Design of Experiment RSM) was used to investigate seven processing factors including: drying and rehydration modalities, supplementation types and biomaterials as drying matrices. The response of adherent and suspension cell lines to the top novel defined protocols included assessment of cytotoxicity, metabolic activity, L/D staining, cell cycle, and functionality/differentiation capacity following rehydration. Finally, two in vivo models were used to assess the dried/rehydrated constructs via the subcutaneous implantation/volumetric muscle loss injury model in SD wildtype rats and C57BL/6 mice. Results: Results in this project provide a comparative profile of plant-based and animal-derived material in terms of thermo-physical, mechanical, permeability and biocompatibility characteristics. Systematic experimentation enabled optimisation of desiccation-rehydration process maximising cell viability and functionality in two cell lines with different growing characteristics. Moreover, novel supplementation and cell protective matrix regimens were demonstrated. Finally, in ovo and in vivo models offered valuable insights on tissue responses with promising contribution of desiccated/rehydrated constructs to normal angiogenesis, M1-to-M2 polarisation and tissue regeneration. Conclusion: Taken together, results demonstrate the feasibility of a desiccation protocol which offers a cold-chain free, more accessible and cost-effective technique for the long-term storage and delivery of cells used in a variety of biomedical and therapeutical applications.

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