Traditional meat production has major sustainability and ethical concerns. Cultivated meat helps to address these concerns by reducing the need for mass animal farming. Muscle progenitor cells (MPCs) harvested from skeletal muscle are a promising cell source for cultivated meat. While various protocols have been developed for MPC isolation, which protocol is best suited for the cultivated meat industry requires further investigation. Therefore, the purpose of this thesis was to optimize the MPC isolation technique to produce a pure myogenic cell population and provide the cultivated meat industry with standardized procedures for production. For these proof-of-concept experiments, skeletal muscles harvested from the hindlimb muscles of mice were used. Cells were isolated from the harvested muscle then subjected to one of three protocols for MPC enrichment: pre-plating, ice-cold treatment (ICT), or fluorescence activated cell sorting (FACS). The pre-plating and ICT protocols resulted in impure cell populations with few MPCs after one week in culture. Therefore, FACS using two cell-surface markers, NCAM and CD34, was employed as a more specific method for MPC sorting. CD34+NCAM1- cells grew quickly, however, unwanted cell types remained following FACS. In contrast, CD34+NCAM1+ cells had a consistent small, rounded shape and slow proliferation rate. These cells remained viable in culture for several months and had high Pax7 expression, indicating they were a pure population of myogenic cells. CD34+NCAM1+ cells maintained their capacity to differentiate after culturing for an extended period, demonstrating their potential use for cultivated meat production. The results of this study provide a better understanding of the differences between previously published MPC isolation techniques. Future studies will investigate the potential for CD34+NCAM1+ cells to be grown on a larger scale. These experiments provide insight into MPC populations that may exist in livestock species and will help to streamline the early stages of cultivated meat production. / Thesis / Master of Science (MSc) / Traditional meat production is associated with numerous challenges including animal welfare concerns, human health concerns, and harmful environmental consequences. The global population is predicted to reach 9.7 billion by 2050, emphasizing the importance of alternative food sources to meet the increased food demand. Cultivated meat is a promising new protein source, with the intended purpose of providing a sustainable food source with reduced ethical concerns compared to conventional meat. While there are several challenges to overcome throughout the production process, a major consideration in the early stages of cultivated meat production is cell sourcing. Muscle cells harvested from a tissue biopsy are one proposed starting cell source which has the potential to make up most of the tissue in cultivated meat products. This thesis aimed to improve upon previously published protocols used for muscle cell isolation and provide an optimized cell population for use in cultivated meat production. The cell sorting protocol described in this thesis provides a highly efficient technique for muscle cell purification and long-term growth. The resulting cell population has many characteristics that are pertinent to cultivated meat and may advance the early stages of production.
| Identifer | oai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/28829 |
| Date | January 2023 |
| Creators | Steele, Alexandra P |
| Contributors | Hawke, Thomas J, Medical Sciences |
| Source Sets | McMaster University |
| Language | English |
| Detected Language | English |
| Type | Thesis |
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