Abstract: A new kind of polyurethane (PU) hydrogels have been prepared based on 100% renewably-sourced poly(trimethylene ether) glycol (PO3G). The formed PU hydrogels could meet the requirement of sustainability and development of polyurethane materials, and also broaden the application of 100% renewably-sourced PO3G. A series of PU hydrogels with different PO3G mass fractions were synthesized by a two-step process. Firstly, the reaction of PO3G and poly(ethylene glycol) (PEG) with 1, 6-diisocyanatohexane (HDI) produced -NCO groups terminated prepolymer, which reacted with 1, 4-butanediol further and provided the PU hydrogel. The effects of PO3G content on the structure and properties of PU hydrogels were studied by Fourier Transform Infrared spectrometer (FT-IR), Differential Scanning Calorimeter (DSC), Dynamic Thermomechanical Analysis (DMA), rheometer, Scanning Electron Microscope (SEM) and measurement of mechanical properties. The results showed that with the decrease of PO3G mass fraction from 80% to 0, the glass transition temperature of PU increased from -55℃ to -29℃, and the crystallinability of soft segments in the PU was enhanced. The swelling ratio of PU increased from 5% to 394%, and the primary swelling behavior of PU hydrogels transited from the Fickian to non-Fickian diffusion. Both the storage modulus and loss modulus of PU hydrogels decreased due to the fact that the water absorption ratio of PU increased when PO3G mass fraction decreased. However, the tensile strength of PUs and PU hydrogels increased significantly with the increase of PO3G content in the PU.