The impact of long-term culture on primary bovine myoblasts : implications for cultivated meat production and allergenicity
(2025) Trlin, Hamish John Francis
With global demand for meat products growing and traditional methods of production nearing their limits, alternative methods of meat production are required. While cultivated meat is a promising alternative to traditional meat, information about the impact of long-term culture on cultivated cells is lacking. Here, I compare the use of high impact methods of stem cell isolation and enrichment in cultivated meat research and product development to alternative low impact methods, define the impacts of isolation and long-term cell culture on the allergenicity of cultivated cells, and determine the broader impact of long-term cell culture on protein abundance and modification in cultivated cells.
Chapter 2 demonstrates that enzymatic digestion and fluorescence-activated cell sorting remain the preferred methods for myoblast isolation and enrichment. While whole tissue explant isolations and collagen pre-plating enrichment have theoretical benefits as low impact alternatives, these methods were unable to produce sufficiently pure populations of myoblasts for use in long-term cell culture. Chapter 3 then reveals differences in the relative abundance or detection of allergens in native and cultured cells, demonstrating why allergenic equivalence should not be assumed. Significant reductions in the abundance of major protein allergens Bos d 6 and Bos d 13 suggest the allergenic potential of cultivated beef meat may be reduced compared to traditional beef meat. However, an increased detection α-Gal in cultured cells, alongside an increased abundance of currently insignificant protein allergens ENO1, ATP5PO, and COL1A2, suggests that allergenicity may be increased. Finally, Chapter 4 identifies significant, ongoing differences in the proteomes of primary bovine myoblast proteomes throughout the duration of long-term culture. More than 70% of proteins are shown to be significantly differently abundant following short or long-term culture, accompanied by changes in the general patterns of glycan modification. These results clearly demonstrate that culture duration has a significant effect on primary stem cell populations.
This thesis provides evidence that allergens are differently abundant in native and cultivated cells, calling to question the present assumption of allergenic equivalence between traditional and cultivated meats. While the clinical significance of these results have yet to be determined, they highlight a need for the optimisation of myoblast isolation and culture to reduce differences between native and cultured cells. Minimising these differences is likely to improve the characteristics of primary cultivated cells, thus cultivated meat, while reducing allergenic potential.