Abstract
Cultivated meat provides a sustainable and ethical alternative to traditional livestock production, addressing global challenges in meat demand and environmental impact. Fat is recognized as an essential component of meat products, but strategies for culturing animal fat precursors still have major limitations, such as the need for a serum-containing medium and inefficient adipogenic cell maturation. In this study, a serum-free medium (SFM) formulation for the cultivation of bovine adipose-derived stem cells (bASC) was first validated by comparison with serum-containing media (FBS). After a 14-day induction period, the SFM increased bASC proliferation by 20% and triglyceride accumulation by 66%. When the SFM was applied to a three-dimensional (3D) spheroid culture system, it resulted in up to 34% higher triglyceride accumulation compared to 2D cultures. These results were accompanied by a significant increase in the expression of key adipogenic markers and lipid droplet formation, suggesting a synergistic potential of SFM and 3D spheroid strategies for bASC culture. Overall, the results of this study provide a scalable and sustainable platform for cultured fat production and are an important step towards overcoming some of the challenges in cultured meat production.