Small molecule cocktail for the serum-free Adipogenic induction of adipose-derived stem cells from Larimichthys crocea: Towards the efficient production of cell-cultured fish fat

This study aims to develop a safe, efficient, and serum-free induction platform for the adipogenic differentiation of adipose-derived stem cells (ASCs) from Larimichthys crocea , to advance the production of cultivated fat for cellular agriculture. Through systematic screening of plant-derived small molecules, bavachinin and dehydroabietic acid were identified as core inducing components, with their concentrations and exogenous lipid supplementation strategies optimised. A serum substitute culture system (SS) was successfully established, comprising growth factors, antioxidants, and carrier proteins, which effectively supports ASCs adipogenesis. Further investigation under three-dimensional culture on gelatin-based porous microcarriers revealed that lower stiffness (10 kPa) enhanced lipid accumulation and adipogenic gene expression. Transcriptomic and metabolomic analyses indicated that the induction system activates the PPARγ pathway and promotes lipid synthesis and accumulation. This work provides a robust chemical biology strategy and theoretical foundation for the scalable production of cultivated fish adipose tissue.