Caffeine inhibits adipogenic differentiation of primary adipose-derived stem cells and bone marrow stromal cells

- Caffeine suppressed adipogenic differentiation of ADSCs and M2-10B4 cells.
- Caffeine could reduce the accumulation of lipid droplets, expression of adipogenic genes, and formation of adipocyte.
- Caffeine effectively inhibited adipogenic differentiation through downregulating adipogenesis-related genes and cytokines.
- Low dose of caffeine inhibited adipogenesis, whereas high dose of caffeine had cytotoxicity to adipose-derived stem cells.

Caffeine consumption has been related to loss of body weight and modulates lipid metabolism. However, impacts of caffeine on adipogenic differentiation have not been well determined yet. The present study evaluated the effects of caffeine on adipogenesis using primary rat adipose-derived stem cells (ADSCs) and a mouse bone marrow stromal cell line (M2-10B4) in vitro. ADSCs and M2-10B4 were continuously exposed to caffeine (0.1-1 mM) during adipogenic differentiation for 7 and 12 days, respectively. Oil red O and Nile red staining showed that caffeine reduced lipid droplet and adipocyte levels in both cell types. In addition, Nile red staining and FACScan flow cytometry showed that caffeine dose-dependently decreased adipocyte differentiation from 20% to 50% of the control ADSCs and M2-10B4 cells. Caffeine decreased the expression of adipogenesis-related genes including peroxisome proliferator-activated receptor-γ, CCAAT/enhancer-binding protein-α, adipocyte lipid binding protein, lipoprotein lipase, leptin, and TNFα in a dose-dependent manner. Rather, low concentration of caffeine (0.1 mM) significantly increased IL-6 expression, but unexpectedly inhibited that at a concentration more than 0.3 mM. Taken together, caffeine was able to effectively inhibit adipogenic differentiation of ADSCs and M2-10B4 cells partly through its inhibition of adipogenesis-related factors.