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Single-cell analysis of bovine muscle-derived cell types for cultured meat production

ABSTRACT: Single-cell analysis of bovine muscle-derived cell types for cultured meat production

PROVIDER: PRJNA870300 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21101600_1.fastq.gz	Fastqsanger.gz
	SRR21101600_2.fastq.gz	Fastqsanger.gz
	SRR21101601_1.fastq.gz	Fastqsanger.gz
	SRR21101601_2.fastq.gz	Fastqsanger.gz
	SRR21101602_1.fastq.gz	Fastqsanger.gz

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Single-cell analysis of bovine muscle-derived cell types for cultured meat production

Project description:Cultured' or 'cultivated' 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 cell types, including muscle fibres, satellite cells (SCs) and fibro-adipogenic progenitors (FAPs), and accurate recreation of the entire tissue thus requires the identification, purification, proliferation and characterisation of a broad range of cell types. Here, we use a single-cell RNA sequencing approach to characterise transcriptional heterogeneity within bovine muscle and derived in vitro 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 favour the growth of SCs in the context of serum-free medium. Combining RNA velocities computed in silico with functional data, we characterise dynamic subpopulations and transitions between active, quiescent and committed substates of SCs, and demonstrate methods for modulation of these states during long-term proliferative cultures. This work represents an important resource for advancing our knowledge of bovine skeletal muscle biology, and its application in the development of cultured meat technologies.

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Single-cell analysis of bovine muscle-derived cell types for cultured meat production.

Project 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.

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Muscle-derived fibro-adipogenic progenitor cells for production of cultured bovine adipose tissue

Project description:Cultured meat is an emergent technology with the potential for significant environmental and animal welfare benefits. Accurate mimicry of traditional meat requires fat tissue; a key contributor to both the flavour and texture of meat. Here, we show that fibro-adipogenic progenitor cells (FAPs) are present in bovine muscle, and are transcriptionally and immunophenotypically distinct from satellite cells. These two cell types can be purified from a single muscle sample using a simple fluorescence-activated cell sorting (FACS) strategy. FAPs demonstrate high levels of adipogenic potential, as measured by gene expression changes and lipid accumulation, and can be proliferated for a large number of population doublings, demonstrating their suitability for a scalable cultured meat production process. Crucially, FAPs reach a mature level of adipogenic differentiation in three-dimensional, edible hydrogels. The resultant tissue accurately mimics traditional beef fat in terms of lipid profile and taste, and FAPs thus represent a promising candidate cell type for the production of cultured fat.

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A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation [SF_Diff]

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A serum-free media formulation for cultured meat production supports bovine satellite cell differentiation in the absence of serum starvation

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