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Biology subjects

Spaans, S.

Publications and source records attributed to Spaans, S..

3 recordsLinked to original sources

Peptide screening enables optimised biofunctional hydrogels for cultivated meat tissue engineering

Cultivated meat is an emerging biotechnology that aims to produce edible tissues in an ethical and sustainable manner. However, the recreation of skeletal muscle tissue that replicates the protein composition and sensory characteristics of traditional meat is a major challenge. Skeletal muscle tissue engineering requires non-animal-based scaffolds which are inexpensive and food-safe, while meeting specific mechanical requirements with respect to viscosity, stress-relaxation and stiffness. While many of these characteristics can be fulfilled by alginate-based biomaterials, a key limitation of alginate is its lack of intrinsic attachment sites for animal cells, preventing efficient adhesion, differentiation and tissue formation. Here, we established a screening platform to evaluate extracellular matrix (ECM)-mimicking peptides as functionalisations of alginate scaffolds in 2D. Our platform enables high-throughput assessment of cell/peptide interactions, serving as a predictive tool for 3D tissue constructs. Our screen identified two RGD-containing sequences (vitronectin- and fibronectin-mimicking peptides) as most effective in promoting attachment and myogenic fusion of bovine satellite cells. Notably, these peptides outperformed more complex mixtures containing up to seven different ECM-mimicking peptides. Our findings provide a streamlined approach for optimising biomaterial functionalisations for cultivated meat applications, and lay the groundwork for future advancements in scalable, sustainable skeletal muscle tissue engineering.

bioengineering↗

Short-stranded zein fibers for muscle tissue engineering in alginate-based hydrogels

Cultivated meat is a nascent technology that aims to produce an environmentally and animal-friendly alternative to conventional meat. Producing skeletal muscle tissue in an animal-free system allowing for high levels of myofusion and maturation is important for the nutritional and sensorial value of cultivated meat. Alginate is an attractive biomaterial to support muscle formation as it is food-safe, sustainable, cheap, and can be cross-linked using non-toxic methods. Although alginate can be functionalized to promote cell attachment, limitations in its mechanical properties, including form, viscosity and stress relaxation, hinder cellular capacity for myogenic differentiation and maturation in alginate-based hydrogels. Here, we show that the addition of electrospun short-stranded zein fibers increased hydrogel degradation, resulting in faster compaction, improved cell-gel interaction and enhanced alignment of bovine muscle precursor cells. We conclude that fiber-hydrogel composites are a promising approach to support optimal formation of 3D constructs, by improving tissue stability and thus prolonging culture duration. Together, this improves muscle-related protein content by facilitating myogenic differentiation and priming muscle organoids for maturation.

bioengineering↗

Optimisation of cell fate determination for cultured muscle differentiation

Production of cultured meat requires defined medium formulations for the robust differentiation of myogenic cells into mature skeletal muscle fibers in vitro. Whilst such formulations can drive myogenic differentiation to an extent similar to serum-starvation based protocols, these cultures are invariably heterogeneous in nature, with a significant proportion of cells not participating in myofusion, limiting maturation of the muscle. Here, we use RNA sequencing to characterise this heterogeneity at single-nucleus resolution, identifying distinct cellular subpopulations, including proliferative cells that fail to exit the cell cycle, and reserve cells that do not commit to myogenic differentiation. We show that the ERK, NOTCH and RXR pathways act during the first stages of myogenic cell fate determination, and by targeting these pathways, cell cycle exit can be promoted whilst abrogating reserve cell formation. Under these improved culture conditions, fusion indices close to 100% can be robustly obtained in 2D culture. Finally, we demonstrate that this translates to higher levels of myotube formation and muscle protein accumulation in animal component-free bioartificial muscle constructs, providing proof of principle for the generation of highly differentiated cultured muscle with excellent mimicry to traditional muscle.

cell biology↗