Single-cell analysis of bovine muscle-derived cell types for cultured meat production
Cultured meat technologies leverage the proliferation and differentiation of animal-derived stem cells ex vivo to produce edible tissues for human consumption in a sustainable fashion. However, skeletal muscle is a dynamic and highly complex tissue, involving the interplay of numerous mono- and mult...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-09-01
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| Series: | Frontiers in Nutrition |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fnut.2023.1212196/full |
| _version_ | 1797687156903247872 |
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| author | Tobias Messmer Tobias Messmer Richard G. J. Dohmen Richard G. J. Dohmen Lieke Schaeken Lea Melzener Lea Melzener Rui Hueber Mary Godec Carin Didoss Mark J. Post Mark J. Post Joshua E. Flack |
| author_facet | Tobias Messmer Tobias Messmer Richard G. J. Dohmen Richard G. J. Dohmen Lieke Schaeken Lea Melzener Lea Melzener Rui Hueber Mary Godec Carin Didoss Mark J. Post Mark J. Post Joshua E. Flack |
| author_sort | Tobias Messmer |
| collection | DOAJ |
| description | Cultured meat technologies leverage the proliferation and differentiation of animal-derived stem cells ex vivo to produce edible tissues for human consumption in a sustainable fashion. However, skeletal muscle is a dynamic and highly complex tissue, involving the interplay of numerous mono- and multinucleated cells, including muscle fibers, satellite cells (SCs) and fibro-adipogenic progenitors (FAPs), and recreation of the tissue in vitro thus requires the characterization and manipulation of a broad range of cell types. Here, we use a single-cell RNA sequencing approach to characterize cellular heterogeneity within bovine muscle and muscle-derived cell cultures over time. Using this data, we identify numerous distinct cell types, and develop robust protocols for the easy purification and proliferation of several of these populations. We note overgrowth of undesirable cell types within heterogeneous proliferative cultures as a barrier to efficient cultured meat production, and use transcriptomics to identify conditions that favor the growth of SCs in the context of serum-free medium. Combining RNA velocities computed in silico with time-resolved flow cytometric analysis, we characterize dynamic subpopulations and transitions between active, quiescent, and committed states of SCs, and demonstrate methods for modulation of these states during long-term proliferative cultures. This work provides an important reference for advancing our knowledge of bovine skeletal muscle biology, and its application in the development of cultured meat technologies. |
| first_indexed | 2024-03-12T01:14:14Z |
| format | Article |
| id | doaj.art-f1fe53e87caf4ed4bb17899aca618d9c |
| institution | Directory Open Access Journal |
| issn | 2296-861X |
| language | English |
| last_indexed | 2024-03-12T01:14:14Z |
| publishDate | 2023-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Nutrition |
| spelling | doaj.art-f1fe53e87caf4ed4bb17899aca618d9c2023-09-13T21:53:26ZengFrontiers Media S.A.Frontiers in Nutrition2296-861X2023-09-011010.3389/fnut.2023.12121961212196Single-cell analysis of bovine muscle-derived cell types for cultured meat productionTobias Messmer0Tobias Messmer1Richard G. J. Dohmen2Richard G. J. Dohmen3Lieke Schaeken4Lea Melzener5Lea Melzener6Rui Hueber7Mary Godec8Carin Didoss9Mark J. Post10Mark J. Post11Joshua E. Flack12Mosa Meat B.V., Maastricht, NetherlandsMaastricht University, Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsMaastricht University, Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsMaastricht University, Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsMaastricht University, Maastricht, NetherlandsMosa Meat B.V., Maastricht, NetherlandsCultured meat technologies leverage the proliferation and differentiation of animal-derived stem cells ex vivo to produce edible tissues for human consumption in a sustainable fashion. However, skeletal muscle is a dynamic and highly complex tissue, involving the interplay of numerous mono- and multinucleated cells, including muscle fibers, satellite cells (SCs) and fibro-adipogenic progenitors (FAPs), and recreation of the tissue in vitro thus requires the characterization and manipulation of a broad range of cell types. Here, we use a single-cell RNA sequencing approach to characterize cellular heterogeneity within bovine muscle and muscle-derived cell cultures over time. Using this data, we identify numerous distinct cell types, and develop robust protocols for the easy purification and proliferation of several of these populations. We note overgrowth of undesirable cell types within heterogeneous proliferative cultures as a barrier to efficient cultured meat production, and use transcriptomics to identify conditions that favor the growth of SCs in the context of serum-free medium. Combining RNA velocities computed in silico with time-resolved flow cytometric analysis, we characterize dynamic subpopulations and transitions between active, quiescent, and committed states of SCs, and demonstrate methods for modulation of these states during long-term proliferative cultures. This work provides an important reference for advancing our knowledge of bovine skeletal muscle biology, and its application in the development of cultured meat technologies.https://www.frontiersin.org/articles/10.3389/fnut.2023.1212196/fullsingle-cell RNA sequencingcultured meatcell purificationmuscle stem cellssatellite cell heterogeneity |
| spellingShingle | Tobias Messmer Tobias Messmer Richard G. J. Dohmen Richard G. J. Dohmen Lieke Schaeken Lea Melzener Lea Melzener Rui Hueber Mary Godec Carin Didoss Mark J. Post Mark J. Post Joshua E. Flack Single-cell analysis of bovine muscle-derived cell types for cultured meat production Frontiers in Nutrition single-cell RNA sequencing cultured meat cell purification muscle stem cells satellite cell heterogeneity |
| title | Single-cell analysis of bovine muscle-derived cell types for cultured meat production |
| title_full | Single-cell analysis of bovine muscle-derived cell types for cultured meat production |
| title_fullStr | Single-cell analysis of bovine muscle-derived cell types for cultured meat production |
| title_full_unstemmed | Single-cell analysis of bovine muscle-derived cell types for cultured meat production |
| title_short | Single-cell analysis of bovine muscle-derived cell types for cultured meat production |
| title_sort | single cell analysis of bovine muscle derived cell types for cultured meat production |
| topic | single-cell RNA sequencing cultured meat cell purification muscle stem cells satellite cell heterogeneity |
| url | https://www.frontiersin.org/articles/10.3389/fnut.2023.1212196/full |
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