Muscle Tissue Maturation using Non-animal Derived Biomaterials-based Scaffolds, for Cultured Meat Development

Background: Clean meat is a promising candidate to cope with the rising meat products demand over the past decades, known to cause environmental, health and ethical problems. For this purpose, tissue engineering techniques are harnessed to create an edible lab grown muscle tissue, made from suitable biomaterials & cell types. The scaffolding materials must meet key requirements to enable cell growth & maturation, and will preferably contain significant nutritional values. Here we aimed to engineer nutritious scaffolds to fabricate an edible bovine muscle tissue.

Methods: 3 types of scaffolds were fabricated and assessed for Bovine Satellite Cells (BSCs) growth and differentiation. All compositions are bio-compatible, while 2 out of the 3 types also possess nutritional values. All were assessed in terms of BSCs seeding efficiency (cell counting), coverage fold change (image analysis), and differentiation (confocal imaging & RT-qPCR). Initial mechanical properties were assessed as well – such as swelling ratio (weight fold-change post liquid absorption).

Results: BSCs attachment & proliferation were successful in all scaffold types. Differentiation was later found to also be evenly successful, as indicated by the immunostaining of Desmin & Myogenin, revealing multi-nucleated myotubes in all types – which was strengthened by RT-qPCR for MyoD & Myogenn gene expression. Swelling ratio was found high for all scaffolds, and could be harnessed for further platform development.

Conclusions: We established fabrication protocols of nutritious scaffolds, capable of supporting BSCs growth and differentiation, as a step towards an edible engineered muscle tissue.