Abstract: OBJECTIVE To explore the effects and molecular mechanisms of hypoxia on hMSCs differentiation. METHODS hMSCs were cultured under normoxic or hypoxic conditions; hMSCs differentiation were obtained by treated with BDM(bone differentiation medium); Calcium deposit was assessed by Alizard Red S dye; The protein levels were analyzed by western blotting. RESULTS Hypoxia can significantly promoted calcium deposit in hMSCs; The expression of LC3 was increased in hMSCs, hypoxia induced calcium deposit was inhibited by autophagy inhibitor 3MA; Hypoxia induced hMSCs calcium deposit was accompanied by inactivation of MAPK and PI3K-AKT-mTOR pathway, inhibition of these pathway could mediate hMSCs calcium deposit by promoted autophagy. CONCLUSION Hypoxia can significantly triger hMSCs differentiation by induced autophagy, inhibition of MAPK and PI3K-AKT-mTOR pathway could contribute to autophagy which could mediate hMSCs differentiation.