Transdifferentiation of fibroblasts into muscle cells to constitute cultured meat with tunable intramuscular fat deposition

eLife, Journal Year: 2024, Volume and Issue: 13

Published: Jan. 22, 2024

Current studies on cultured meat mainly focus the muscle tissue reconstruction in vitro, but lack formation of intramuscular fat, which is a crucial factor determining taste, texture, and nutritional contents. Therefore, incorporating fat into superior value. In this study, we employed myogenic/lipogenic transdifferentiation chicken fibroblasts 3D to produce mass deposit same cells without co-culture or mixture different substances. The immortalized embryonic were implanted hydrogel scaffold, cell proliferation myogenic conducted whole-cut mimics. Compared 2D, grown matrix showed elevated myogenesis collagen production. We further induced deposition transdifferentiated triglyceride content could be manipulated match exceed levels meat. gene expression analysis indicated that both lineage-specific multifunctional signalings contribute generation muscle/fat matrix. Overall, able precisely modulate muscle, extracellular contents according balanced specialized preferences. These findings provide new avenues for customized production with desired can tailored meet diverse demands consumers.

Language: Английский

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MSTN knockout enhances the production of MYOD1-mediated steak-type cultivated meat

Journal of Animal Science and Biotechnology/Journal of animal science and biotechnology, Journal Year: 2025, Volume and Issue: 16(1)

Published: March 11, 2025

Abstract Background As the global population increases, demand for protein sources is expected to increase, driving cell-based cultivated meat. This study aimed enhance productivity of meat through optimization cell source and organization process. Results We engineered fibroblasts into myogenic cells via non-viral introduction MYOD1 gene, avoiding viral methods safety. After confirming stable derivation cells, we combined knockout (KO) MSTN , a negative regulator myogenesis, with -mediated myogenesis improve production. Primary from KO cattle exhibited enhanced potential. Additionally, when tested in immortalized fibroblasts, myostatin treatment reduced -induced two-dimensional cultures, while increased it. To achieve muscle-like alignment, employed digital light processing (DLP)-based three-dimensional (3D) bioprinting organize 3D groove-shaped hydrogels. These bioactive hydrogels supported proliferation significantly improved muscle alignment. Upon differentiation myotubes, demonstrated an ordered particularly which showed highly efficient differentiation. Conclusions The integration genetic modification advanced DLP groove-patterned provides effective strategy producing high-quality, muscle-aligned

Language: Английский

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Fluorescein-based SynNotch adaptors for regulating gene expression responses to diverse extracellular and matrix-based cues

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 20, 2025

Synthetic Notch (SynNotch) receptors function like natural proteins and can be used to install customized sense-and-respond capabilities into mammalian cells. Here, we introduce an adaptor-based strategy for regulating SynNotch activity via fluorescein isomers analogs. Using optimized fluorescein-binding receptor, describe ways chemically control signaling, including approach based on a bio-orthogonal chemical ligation spatially controllable the photo-patterned uncaging of o-nitrobenzyl-caged conjugate. We further show that fluorescein-conjugated extracellular matrix (ECM)-binding peptides regulate depending folding state collagen-based ECM networks. To demonstrate utility these tools, apply them activate dose-dependent gene expression responses induce myogenic-like phenotypes in multipotent fibroblasts with spatiotemporal microenvironmental control. Overall, as versatile tool transcriptional ligands clinically-approved dye. endow cells custom capabilities. authors controlling diverse stimuli fluorescein-conjugates.

Language: Английский

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LncRNA SNHG1 regulates muscle stem cells fate through Wnt/β‐catenin pathway

Developmental Dynamics, Journal Year: 2025, Volume and Issue: unknown

Published: March 21, 2025

Abstract Background Skeletal muscle stem cells (MuSCs) played an important role in maintaining the proper function of tissues. In adults, they normally remained a quiescent state and activated upon stimulation to undergo self‐renewal or myogenic differentiation. This process was complexly regulated by cytokines, molecular mechanisms that promoted MuSCs activation largely unknown. Results Here, we analyzed transcriptome data from different stimuli using weighted gene co‐expression network analysis (WGCNA) identified key long non‐coding RNA SNHG1 (lncSNHG1), which promotes transition MuSCs. Overexpression lncSNHG1 able promote proliferation differentiation MuSCs, whereas knockdown resulted opposite results. Mechanistically, disruption Wnt/β‐catenin pathway blocked quiescence exit induced lncSNHG1. Conclusions We conclude is factor cell through pathway.

Language: Английский

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3D Bioprinting of Cultivated Meat Followed by the Development of a Fine-Tuned YOLO Model for the Detection and Counting of Lipoblasts, Fibroblasts, and Myogenic Cells

Frontiers in Bioscience-Landmark, Journal Year: 2025, Volume and Issue: 30(3)

Published: March 18, 2025

Background: Cultured meat holds significant potential as a pivotal solution for producing safe, sustainable, and high-quality protein to meet the growing demands of global population. However, scaling this technology requires innovative bioengineering approaches integrated with software methods assess growth cell cultures. This study aims develop 3D printing hybrid product subsequently design finely tuned You Only Look Once (YOLO) model detecting counting lipoblasts, fibroblasts, myogenic cells. Methods: Cultures multipotent mesenchymal stem cells (MMSCs) fibroblasts were obtained from domestic rabbit Oryctolagus cuniculus domesticus. Standard protocols employed induce adipogenic differentiation MMSCs. Fibroblasts isolated skin biopsy samples. The process utilized bioinks. engineering approach involved development unique print head into printer. Confocal transmission electron microscopy within construct was performed. A dataset digital images cells, created. Four models based on YOLOv8-seg architecture trained annotated images, implemented in Telegram bot. Results: Stable cultures obtained. 3D-printed tissue constructs composed sodium alginate, sunflower successfully fabricated. printer assembled. confirmed viability construct. Ultrastructural analysis revealed dense intercellular contacts high metabolic activity. resulting replicated organoleptic structural properties natural meat. In IT segment, single-class lipoblasts achieved metrics recall 85%, precision 77%, mean Average Precision at IoU threshold 0.50 (mAP50) 79%, which improved multiclass 92%, mAP50 81%. bot capable different types. Conclusions: achieved. Detailed microscopic demonstrated activity polymerized alginate hydrogel. engineered presents alternative Additionally, neural network models, bot, proved effective monitoring culture identifying types across three As result, we developed four YOLOv8 that outperforms model. all exhibited reduced accuracy high-density cultures, where overlapping led undercounting.

Language: Английский

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Differentiated muscle cells of salamander Pleurodeles waltl re-enter the cell cycle

Histochemistry and Cell Biology, Journal Year: 2025, Volume and Issue: 163(1)

Published: April 17, 2025

Language: Английский

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Lipid Droplet‐Derived Biomimetic Nanocarriers for the Enhancement of Porcine Intermuscular Fat Content

Advanced Science, Journal Year: 2025, Volume and Issue: unknown

Published: May 14, 2025

Abstract Intramuscular fat (IMF) content is a critical factor influencing the quality and palatability of pork. However, established breeding nutritional methods lead to systemic deposition. The efficient specific increase IMF in livestock remain challenge. In this study, Lipid droplets (LDs), an organelle cells, are shown be controllable biocompatible carrier that can used content. This study demonstrated isolated LDs stimulate lipid synthesis by encapsulating Oleic Acid (OA), PPARγ ligand. Importantly, artificial (aLDs) effectively substitute LDs. vivo, investigations showed OA‐loading aLDs significantly increased expression synthases such as DGAT2 FASN through activation PPARγ, key pathway for synthesis. Furthermore, on injected side up 35% with minimal effects. development OA‐aLDs promotes deposition demonstrates significant potential drug carriers. exploration LDs‐based carriers provides new theoretical basis their application medical field.

Language: Английский

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Tailoring the taste of cultured meat

eLife, Journal Year: 2024, Volume and Issue: 13

Published: May 30, 2024

A new protocol can customize the flavor of lab-grown meat by controlling level fat deposited between muscle cells.

Language: Английский

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Unlocking the potential of cultivated meat through cell line engineering

iScience, Journal Year: 2024, Volume and Issue: 27(10), P. 110877 - 110877

Published: Sept. 6, 2024

Language: Английский

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Unraveling the Complexity and Advancements of Transdifferentiation Technologies in the Biomedical Field and Their Potential Clinical Relevance

Archivum Immunologiae et Therapiae Experimentalis, Journal Year: 2024, Volume and Issue: 73(1)

Published: Dec. 5, 2024

Abstract Chronic diseases such as cancer, autoimmunity, and organ failure currently depend on conventional pharmaceutical treatment, which may cause detrimental side effects in the long term. In this regard, cell-based therapy has emerged a suitable alternative for treating these chronic diseases. Transdifferentiation technologies have evolved therapeutic that converts one differentiated somatic cell into another phenotype by using transcription factors (TFs), small molecules, or small, single-stranded, non-coding RNA molecules (miRNA). The transdifferentiation techniques rely simple, fast, standardized, versatile protocols with minimal chance of tumorigenicity genotoxicity. However, there are still challenges limitations need to be addressed enhance their clinical translation percentage near future. Taking account, we delineated features strategies used techniques. Then, delved different intermediate states were attained during transdifferentiation. Advancements field tissue engineering, cancer dissected. Furthermore, limitations, challenges, future perspectives outlined review provide whole new picture molecular biology, interdisciplinary research, bioinformatics, artificial intelligence will push frontiers technology further establish avenues biomedical research.

Language: Английский

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