L4-70178 Development of Animal-Free Growth Conditions for Enhanced Skeletal Muscle Cell Proliferation, Differentiation, and Myosin Production
The project aims to develop animal component-free culture conditions for efficient skeletal muscle cell proliferation, differentiation, maturation, and production of active myosin.
Period: 01. 03. 2026 - 28. 02. 2029
Partners: COBIK, University of Ljubljana
Skeletal muscle cells and tissues are essential for a growing range of biotechnological and biomedical applications, including cultivated meat production, regenerative medicine, disease modelling, drug testing, and the production of muscle motor proteins. Traditionally, these cells are cultured in media supplemented with foetal bovine or horse serum. However, serum use raises ethical and safety concerns, introduces batch-to-batch variability, and limits control over culture conditions. Animal component-free (ACF) media can address these limitations, but their composition must be carefully optimized to support not only cell proliferation but also differentiation, maturation, and the formation of functional contractile proteins.
The project will study complementary animal skeletal muscle cell systems comprising satellite cells and myoblasts. Reference growth and differentiation parameters will first be established in conventional serum-containing media. Subsequently, separate ACF formulations will be developed for cell proliferation and differentiation. Commercial serum replacements and formulations reported in scientific literature will be evaluated according to their effects on cell growth, myoblast fusion, myotube formation, expression of muscle-specific markers, and overall cost efficiency.
An important objective is the production of endogenous and recombinant myosin in its active form under serum-free conditions. Recombinant myosin will be produced using differentiated C2C12 cells, while laboratory-grown animal skeletal muscle cells will provide endogenous myosin. The purified proteins will be characterized using biochemical and functional assays. COBIK will mainly lead the development of ACF media while Faculty of Biotechnology, will mainly contribute to primary cell culture development.
The project will provide new tools and methods for sustainable and reproducible muscle cell cultivation. Its results are expected to support cellular agriculture and cultivated meat production while also advancing research in muscle biology, tissue engineering, regenerative medicine, disease modelling, and myosin-based technologies.
The project is cofunded by the Slovenian Research and Innovation Agency ARIS and Tech4Meat d.o.o.
