We aimed to examine the anti-adipogenic effects of mangiferin (xanthone glucoside) and its related mechanism. Human mesenchymal stem cells (hMSCs) were cultured in Dulbecco's modified Eagle medium, after 60% confluence adipocyte differentiation was induced. Differentiation-induced hMSCs were cultured in the presence and absence of 10, 20 and 40 μmol of mangiferin from day 0 to day 10. Adipocyte differentiation and lipid accumulation were significantly decreased in 40 μmol mangiferin-treated groups when compared with the reference drugs (quercetin- and orlistat-treated groups). Using quantitative reverse transcription polymerase chain reaction, we studied the mRNA expression levels of resistin, adipocyte fatty acid-binding protein 2 (aP2), lipoprotein lipase (LPL), peroxisome proliferator-activated receptor (PPAR-γ) and tumor necrosis factor-α, in hMSCs undergoing adipocyte differentiation; treatment with mangiferin attenuated the expression of those adipogenic genes and decreased adipocyte differentiation. Mangiferin significantly inhibited hMSCs to preadipocyte differentiation within the first 2 days of treatment, indicating that the anti-adipogenic effects of mangiferin are achieved through the inhibition of differentiation and maturation. Practical Applications It is well known that various plant derived compounds presents antiobesity effect. However still there is lack of drugs for the arrest or inhibition of adipocyte differentiation from mesenchymal stem cell precursors. hMSCs treated with adipocyte differentiation medium along with mangiferin inhibits preadipocyte differentiation and maturation, via suppressing lipid accumulation. Mangiferin treatment resulted in a significant (p < 0.05) decrease in GPDH and triglyceride levels as well as an increase in LDH activity and attenuated lipid accumulation compared with untreated differentiating preadipocytes. This effect was demonstrated by the observed down-regulation of adipogenesis related genes after hMSCs underwent induced adipocyte differentiation.