Development of a clinically translatable allogenic umbilical cord-derived mesenchymal stem cell conditioned media

Parnell, A. J. (2026) Development of a clinically translatable allogenic umbilical cord-derived mesenchymal stem cell conditioned media. PhD thesis, University of Reading. doi: 10.48683/1926.00129233

Abstract/Summary

Mesenchymal stem cells (MSCs) have featured in the regenerative medical field for decades, with particular focus on their secretomes’ paracrine nature for treating disease or regenerating damage. Conditioned medias and subsequent products using the derivates of MSC secretomes have flooded the space over the last two decades, many of which still use crude research laboratory methods for the generation. Although their therapeutic capacity is well documented, the methods for generating them are often far from what would be required for regulatory approval and clinical manufacture. Here, methods of producing clinically relevant umbilical cord-derived MSC conditioned media were explored. In particular, a novel pelleting approach (UCMSC PEL), was evaluated to reveal potent proliferation and migration enhancing and anti-inflammatory properties. Batch-to-batch and donor to-donor variation was found to be minor. However, the concentration of key components within the secretome and differences in biological outputs compared to an adherent conditioned media (UCMSC ADH), weighed against the UCMSC PEL being a commercially viable product. Thus, UCMSC ADH conditioned media was developed from a research-grade conditioned media to one resembling a clinically compliant, scalable therapeutic candidate with commercial appeal. During the development, the use of bicarbonate buffers or basal medias remained favourable, and scale-up protocols were conceptualised. Finally, mechanisms of action and biological applications of the developed preparation (UCMSC ADH-E) were revealed through in vitro testing to include enhancing proliferation and migration, augmentation of biomolecule activity (metalloproteinases and serine proteases), and potent anti-inflammatory properties against an LPS-induced blood and PBMC assay. Cytokine arrays and gene expression analysis (qPCR) revealed the immunomodulatory nature of UCMSC ADH-E was linked to regulation of NF-κB controlled genes. An in vivo model of muscle regeneration identified anti-fibrotic and anti-haemorrhagic properties, however no acceleration to muscle fibre regeneration was observed, which was later attributed to an anti-differentiation effect from UCMSC ADH-E on myoblast fusion. All in all, this project has identified a clinically relevant UCMSC conditioned media using GMP available material, which is reproducible, scalable, and biologically potent.

Item Type	Thesis (PhD)
URI	https://centaur.reading.ac.uk/id/eprint/129233
Identification Number/DOI	10.48683/1926.00129233
Date on Title Page	September 2025
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