CRISPR screen reveals genes and pathways regulating proliferation in bovine stem cells for cultured meat [Short-Term screen]
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ABSTRACT:
PROVIDER: PRJNA1179950 | ENA |
REPOSITORIES: ENA
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Project description:Cultured meat offers a promising solution to the environmental and ethical challenges of conventional meat production, but faces significant hurdles in scalability and cost-effectiveness. A major bottleneck is the limited proliferation capacity of cell sources such as bovine mesenchymal stem cells (bMSCs). Here, we employ CRISPR knockout screening to identify several key genes and pathways influencing bMSC proliferation. Notably, TP53 and PTEN knockouts emerged as top hits in our pooled CRISPR screen, exhibiting the highest increase in cell abundance over 30 days compared to all other gene knockouts. Subsequent validation using individual knockouts confirmed that both TP53 and PTEN deletion significantly enhanced bMSC proliferation rates relative to non-targeting sgRNA controls. We also identified targets in mitochondrial pyruvate carrier and SMAD signaling pathways that may improve MSC proliferation and differentiation. These findings demonstrate CRISPR technology's potential to enhance stem cell growth for more efficient, cost-effective, and scalable cultured meat production.
2025-08-27 | GSE280676 | GEO
Project description:Cultured meat offers a promising solution to the environmental and ethical challenges of conventional meat production, but faces significant hurdles in scalability and cost-effectiveness. A major bottleneck is the limited proliferation capacity of cell sources such as bovine mesenchymal stem cells (bMSCs). Here, we employ CRISPR knockout screening to identify several key genes and pathways influencing bMSC proliferation. Notably, TP53 and PTEN knockouts emerged as top hits in our pooled CRISPR screen, exhibiting the highest increase in cell abundance over 30 days compared to all other gene knockouts. Subsequent validation using individual knockouts confirmed that both TP53 and PTEN deletion significantly enhanced bMSC proliferation rates relative to non-targeting sgRNA controls. We also identified targets in mitochondrial pyruvate carrier and SMAD signaling pathways that may improve MSC proliferation and differentiation. These findings demonstrate CRISPR technology's potential to enhance stem cell growth for more efficient, cost-effective, and scalable cultured meat production.
2025-08-27 | GSE280666 | GEO
Project description:Cultured meat offers a promising solution to the environmental and ethical challenges of conventional meat production, but faces significant hurdles in scalability and cost-effectiveness. A major bottleneck is the limited proliferation capacity of cell sources such as bovine mesenchymal stem cells (bMSCs). Here, we employ CRISPR knockout screening to identify several key genes and pathways influencing bMSC proliferation. Notably, TP53 and PTEN knockouts emerged as top hits in our pooled CRISPR screen, exhibiting the highest increase in cell abundance over 30 days compared to all other gene knockouts. Subsequent validation using individual knockouts confirmed that both TP53 and PTEN deletion significantly enhanced bMSC proliferation rates relative to non-targeting sgRNA controls. We also identified targets in mitochondrial pyruvate carrier and SMAD signaling pathways that may improve MSC proliferation and differentiation. These findings demonstrate CRISPR technology's potential to enhance stem cell growth for more efficient, cost-effective, and scalable cultured meat production.
2025-08-27 | GSE280675 | GEO
Project description:CRISPR screen reveals genes and pathways regulating proliferation in bovine stem cells for cultured meat [Long-Term screen]
| PRJNA1179933 | ENA
Project description:CRISPR screen reveals genes and pathways regulating proliferation in bovine stem cells for cultured meat [RNA-Seq]
| PRJNA1179947 | ENA
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.
2022-09-02 | GSE211428 | GEO
Project description:This project aimed to compare the proteomic profiles between conventional chicken meat and cultured chicken meat produced through cell culture techniques. Proteins were extracted, separated using SDS-PAGE and two-dimensional gel electrophoresis (2-DE), and identified through mass spectrometry. KEGG pathway enrichment and STRING-based protein-protein interaction network analyses were performed to evaluate metabolic characteristics and structural differences between the groups. Conventional meat exhibited a high abundance of glycolytic and muscle contraction-associated proteins, while cultured meat showed elevated expression of proteins involved in stress response and redox regulation. These datasets provide fundamental insights for improving the quality and ensuring the safety of cultured meat products.
2025-08-04 | MSV000098713 | MassIVE
Project description:The use of an animal component-free scaffold to culture muscle cells on is a potential approach for producing (hybrid) cultured meat. However, to develop a plant-based scaffold with optimal mechanical characteristics for muscle cell growth, differentiation and maturation, it is essential to characterize the natural extracellular matrix (ECM) found in conventional meat. Therefore, this research initially involved isolating the ECM by removing muscle cells through 0.5 % SDS decellularization. Subsequently, various structural parameters of both fresh meat (representing the upper limit) and decellularized meat (representing the lower limit) were measured. Effective removal of cells and adequate preservation of the ECM's main structure and components were confirmed through Picogreen assay, proteomic analysis, and determination of glycosaminoglycan (GAG) content. Samples underwent amplitude sweeps, texture analyses and internal structure visualization. The conditions resembling the natural ECM of muscle cells lie between the determined upper and lower limits for each parameter.
2025-07-17 | PXD051447 | Pride
Project description:Cultured meat can provide a sustainable and more ethical alternative to conventional meat. Most of the research in this field has been focused on developing muscle tissue, as it is the main component of meat products, while very few studies address cultured fat tissue, an essential component in the human diet and determinant of meat quality, flavor, juiciness, and tenderness. Here, we engineered bovine fat tissue for cultured meat and incorporated it within engineered bovine muscle tissue. Mesenchymal stem cells (MSCs) were derived from bovine adipose tissue and exhibited the typical phenotypic profile of adipose-derived MSCs. MSC adipogenic differentiation and maturation within alginate-based three-dimensional constructs were optimized to yield a fat-rich edible engineered tissue. Subsequently, a marble-like construct, composed of engineered bovine adipose and muscle tissues, was fabricated, mimicking inter- and intra-muscular fat structures.
| S-EPMC9452530 | biostudies-literature
Project description:Livestock farming and conventional meat production pose significant environmental, health, and animal welfare challenges. In seeking sustainable alternative solutions, cultivated meat technology typically utilizes differentiation of myogenic progenitor cells (MPCs) into muscle cells for in vitro meat production. However, understanding the molecular determinants governing MPC differentiation into muscle cells, and the potential enhancement of this process through modulation of signaling pathways, remains limited. Herein, we characterized the molecular landscape associated with bovine MPC differentiation in vitro by employing multiomics, and explored its augmentation by small molecules, together leading to identification of media that enhanced myogenic differentiation compared with conventional methods in both 2D cultures and tissue‐engineered 3D skeletal muscle constructs. Through bulk and single‐cell transcriptomics and proteomics, we compared conventional and enhanced differentiation media, demonstrating that the enhanced media gave rise to unique progenitor‐like cell populations, while simultaneously promoting differentiation into myocytes and contractile myotubes expressing a wide array of myogenic markers that more closely resemble bovine muscle cells in vivo. The improved method for promoting myogenic differentiation in 2D and 3D formats, together with the corresponding molecular roadmap, may prove valuable for cultivated meat applications.
2025-07-25 | GSE262675 | GEO