Although accumulating evidences indicate that miRNA emerge as a vital player in cell growth, development, and differentiation, how they contribute to the process of adipocyte differentiation remains elusive. In the present study, we revealed that the expression level of miR-210 was dramatically upregulated during 3T3-L1 adipogenesis. Ectopic introduction of miR-210 into 3T3-L1 cells promoted terminal differentiation as well as the expression of adipogenic markers. MTT assay showed that miR-210 significantly inhibited cell proliferation whereas the BrdU incorporation assay and flow cytometry analysis showed that miR-210 did not impair G1/S phase transition. Further experiments demonstrated that enhanced expression of miR-210 in 3T3-L1 cells provoked adipocyte differentiation via activation of PI3K/Akt pathway by targeting SHIP1, a negative regulator of PI3K/Akt pathway. Moreover, blockade of endogenous miR-210 during adipogenesis significantly repressed adipocyte differentiation. In summary, we have identified miR-210 as an important positive regulator in adipocyte differentiation through the activation of PI3K/Akt pathway. J. Cell. Biochem. 114: 2699–2707, 2013. © 2013 Wiley Periodicals, Inc.