Exploring the potential of subcutaneous and intramuscular adipose tissue as cell sources for cellular agriculture on decellularized vegetable scaffolds.

Cellular agriculture offers a promising solution to the growing demand for alternative proteins. However, it faces challenges, including the selection of an optimal cell source and the co-culturing of different cell types. Adipose tissue (AT) is crucial for meat sensory properties, making its inclusion in cultivated meat essential. Efficient cultured fat production requires a suitable cell source, a key aspect in cellular agriculture. This study aimed to characterize and compare AT Mesenchymal Stromal Cells (AT-MSCs) from different adipose sources (subcutaneous–SAT, intramuscular–IMAT) for cultured meat applications. SAT and IMAT samples from 120kg pigs were collected and compared for: i) tissue morphology (adipocytes area, density, stromal fraction%); ii) derived AT-MSCs potential: cell proliferation (Doubling Time-DT, Colony Forming Unit-CFU), mesenchymal and hematopoietic markers expression, trilineage differentiation (adipogenic, osteogenic, chondrogenic) (International Society for Cellular Therapy criteria). Additionally, preliminary studies were conducted on SAT-MSCs cultured on decellularized cabbage scaffolds to assess 3D adipogenic differentiation. Results showed that SAT had bigger adipocytes than IMAT (p<0.05), while no differences were observed in cell density and stroma%. The characterization and differentiation assays confirmed the stromal potential of cells from both sources, with no significant differences in the parameters. Furthermore, preliminary results indicate that SAT-MSCs cultured on the decellularized cabbage scaffold tended to aggregate and differentiate into adipogenic spheroids, which may better mimic conventional fat. These findings indicate that both fat depots are promising cell sources for cellular agriculture, and that decellularized vegetable scaffolds may support 3D adipogenic differentiation. Further research is needed to validate this approach for cultured fat production.