Food-grade fungal pellets as edible scaffolds for bovine stem cell expansion

Abstract

The scalability of cultivated meat production depends on cost-effective, edible scaffolds that support attachment, proliferation and differentiation of adherent cells whilst meeting food safety and sensory requirements. However, most existing microcarriers are synthetic or of animal-origin, limiting their compatibility with food applications, increasing downstream processing costs, and raising ethical and environmental concerns associated with animal use. For the first time, we present mycelia-based microcarriers derived from food-grade Penicillium strains used in cheese production, as scaffolds for cultivated meat. Eight strains, including novel variants developed through non-GMO techniques (sexual breeding and ultraviolet mutagenesis), were screened for cytotoxicity using bovine adipose-derived stem cells. Out of these, four strains (P. camemberti Myc1; P. roqueforti Myc2, Myc3 and Myc4) were selected for further evaluation based on non-cytotoxic behaviour, ease of handling and pellet size comparable to commercial microcarriers. Morphological characterisation revealed that these strains form highly porous, fibrous pellets with estimated specific surface areas of approximately 4400–5100 cm2/g, providing a favourable architecture for cell growth. All four microcarriers supported strong initial cell attachment, meeting or exceeding industry benchmarks for mesenchymal stem cells in both serum-containing and animal-free media. Growth kinetics diverged between strains, with Myc3 and Myc4 displaying the highest growth rates (≥2-fold increase; μ ≈ 0.015 h−1), and doubling times of 47–48 h. These findings highlight the great promise of fungal pellets for the development of edible scaffolds for cultivated meat production, helping to address a central bottleneck in bringing affordable, high-quality protein to consumers.