Cultured fat as an ingredient for in vitro meat: investigating edible supports for adipogenic differentiation.

The global population is expected to increase, intensifying the demand for alternative and sustainable proteins. Cultured meat represents a possible solution to address this need. Fat plays a crucial role in sensory and nutritional qualities of meat, influencing flavor, texture, and consumer acceptance. However, replicating the structure of conventional meat in vitro remains challenging. This study focuses on cultured fat as a key component of in vitro meat production and aims to develop a preliminary model using 3T3-L1 murine preadipocytes cultured on edible 3D scaffolds, serving as a foundation for future applications with mammalian cells. Two edible supports were investigated: decellularized cabbage leaves and plant-derived chitosan hydrogel. Biocompatibility was evaluated through indirect (EN_ISO_10993-12) and direct cytotoxicity assays, including Alamar Blue and Live/Dead assays at multiple time points. These assays were essential to confirm that scaffolds supported 3T3-L1 cells' growth and viability. Oil Red O staining (OROs) was performed at the end of the adipogenic differentiation process to reveal lipid accumulation, a hallmark of successful fat cell differentiation. Both scaffolds resulted in non-cytotoxic, with cell viability above the lower limit of 70% across all tested time points. Direct cytotoxicity assays confirmed the maintenance of cell viability throughout the culture period. Additionally, both culture supports effectively promoted cell growth and 3D aggregation. OROs revealed lipid accumulation in cells cultured on both supports, indicating successful adipogenic differentiation. In conclusion, these findings indicate that both decellularized cabbage leaf and plant-derived chitosan hydrogel were non-cytotoxic and capable of supporting 3D adipogenic differentiation of 3T3-L1 cells. These supports show potential for cultured fat generation and represent a step forward in cellular agriculture. Future phases of the project will focus on transitioning from murine cell lines to primary mammalian cells, with the ultimate goal of developing adipose tissue for applications in cultured meat production.