Lacto-N-neotetraose (LNnT), an immunomodulatory glycan found on the surface of helminth parasites and in human breast milk has been investigated for its anti-inflammatory and immunosuppressive mechanisms. In this study, we investigated the effects of LNnT-dextran conjugate on the induction of beiging, inhibition of adipogenesis, and the stimulation of lipolysis using 3T3-L1 adipocytes. 3T3-L1 pre-adipocytes were matured using standard differentiation protocol and then treated with 40µg/ml LNnT for 3-24hrs. Treatment of mature 3T3-L1 adipocytes with LNnT induced the beiging of white adipocytes as demonstrated by the enhanced expression of TBX1, TMEM26, and PPARγ. LNnT increased both the expression of uncoupling protein 1 (UCP1) and oxygen consumption rate demonstrating the activation of thermogenesis. In addition, LNnT stimulated an increase in mitochondrial biogenesis as evidenced by an increase in mitochondrial content and PGC-1α expression. LNnT also stimulated lipolysis indicated by increased levels of glycerol release. Treatment with LNnT did not stimulate the inhibition of adipogenesis. Statistical analysis was conducted using one-way analysis of variance (ANOVA) on GraphPad Prism software, followed by Tukey’s post hoc tests. These results provide evidence that LNnT demonstrates multifaceted anti-obesity effects and could therefore have potential therapeutic implications on pro-inflammatory diseases such as diabetes and obesity.