UNLABELLED: Beige adipocytes, crucial for thermogenesis, offer a potential therapeutic strategy for obesity. This study investigated the anti-obesity effects of nonanoic acid (NoA), medium-chain fatty acids, and cholecystokinin-8 (CCK-8) on beige adipogenesis in C3H10T1/2 mesenchymal stem cells (C3H10T1/2 MSCs). We observed a significant increase in cholecystokinin B receptor expression in beige adipocytes compared to preadipocytes. The co-treatment with NoA and CCK-8 enhanced beige adipocyte differentiation and lipid accumulation. Moreover, the co-treatment with NoA and CCK-8 upregulated the mRNA expression of thermogenic genes and increased mitochondrial activity more effectively than individual treatment. Specifically, NoA and CCK-8 co-treatment also elevated the protein expression of uncoupling protein 1 and peroxisome proliferator-activated receptor-gamma coactivator-1 alpha. These findings suggest that the additive effect of NoA and CCK-8 promotes the beiging/browning of body fat in beige adipogenesis, potentially serving as an effective approach in the prevention and treatment of obesity and insulin resistance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10068-024-01699-6.