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The extracellular matrix of meat: a characterization towards a 3D scaffold for cultured meat

ABSTRACT: 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.

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ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Muscle

SUBMITTER: BRAY FABRICE

LAB HEAD: fabrice bray

PROVIDER: PXD051447 | Pride | 2025-07-17

REPOSITORIES: Pride

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Project description:Oxygen and carbon dioxide are common protective gases used in modified atmosphere packaging (MAP) of meat. Within the package, they selectively suppress members of the spoilage microbiome, reshaping it to adapted species concomitantly growing upon MAP. Thus, this species must exhibit adaptation mechanisms to withstand the inhibitory effect of carbon dioxide and oxygen, and cope with selective nutrition on MAP meat. In order to uncover these mechanisms, the typical representative meat-spoiling bacteria Brochothrix (B.) thermosphacta TMW2.2101 and four lactic acid bacteria (LAB) Carnobacterium (C.) divergens TMW2.1577, C. maltaromaticum TMW2.1581, Leuconostoc (L.) gelidum subsp. gelidum TMW2.1618 and L. gelidum subsp. gasicomitatum TMW2.1619 were grown in a meat simulation medium under a controlled, sterile environment, aerated constantly with either air, 100%_N2, 30%_CO2/70%_O2 or 30%_CO2/70%_N2. Growth dynamics were monitored and a label-free quantitative mass spectrometric approach was employed to determine changes within the bacterial proteomes in response to the different gas atmospheres. Revealed bacterial tolerance to modified atmospheres (MA) comprise two possible scenarios: Either bacteria were intrinsically adapted to MA, exhibiting no proteomic regulation of enzymes (L. gelidum subsp. gelidum and gasicomitatum) or, tolerance was provided by varying specific metabolic adaptation (B. thermosphacta, C. divergens, C. maltaromaticum). In detail, metabolic adaptation mechanisms to oxygen comprised an enhanced oxidative stress reduction response, adjustment of the pyruvate metabolism and catabolic oxygen consuming reactions. Adaptation to carbon dioxide was characterized by an upregulation of proteins involved in intracellular pH homeostasis, maintenance of osmotic balance and alteration of the fatty acid composition of the cell membrane. We furthermore predict species-specific strategies for different and preferential carbon source utilization enabling a non-competitive coexistence on meat and resulting in a synergistic spoilage. We conclude that a gas atmosphere containing 30%_CO2/70%_O2 has no inhibitory effect on the analyzed prominent meat-spoiling bacteria whereas 30%_CO2/70%_N2 predictively inhibits C. divergens TMW21577 and B. thermosphacta TMW2.2101 but not the other three species. This gives a mechanistically explanation of their acknowledged status as typical spoilage organisms on packaged meats.

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The extracellular matrix of meat: a characterization towards a 3D scaffold for cultured meat

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