Cultivated meat, developed through cell culture technology, is emerging as a promising solution that closely mimics both the flavor and nutrient profiles of conventional meat. One key component that contributes to the flavor of meat is fat content. In this study, bovine adipose-derived stem cells (bADSCs) were cultured for the production of alternative fat in vitro. The expression of mesenchymal stem cell (MSC) markers (CD29, CD73, and CD105) and colony forming efficiency were assessed to characterize bADSCs. bADSCs were differentiated into adipocytes to produce cultivated fat in 2D or 3D culture. The cultivated fat was analyzed by gas chromatography to verify the similarity of the fatty acids of animal-derived fat. Our results show that bADSCs have characteristics of MSC and could differentiate into adipocyte. The ratio of unsaturated fatty acids and saturated fatty acids in cultivated fat and adipose tissue was similar. Adipogenic differentiation of ADSCs using a textured vegetable protein (TVP) scaffold could form the lipid droplets in the TVP. This study demonstrated the establishment of a culture system for the fat production from bADSCs in vitro. The fat produced through bADSCs shows the potential to be used in the composition of hybrid-cultivated meat.