A series of polyurethanes (PUs) containing grafted cholesterol (UA series) and a control series blended with free cholesterol (UB series) were prepared: the spectroscopic, thermal, tensile, shape memory, and low temperature flexibility properties of these series were compared with those of unmodified PU. For both the UA and UB series, the soft segment melting temperature (T m ) was not affected by the cholesterol content. Differential scanning calorimetry (DSC) results showed that the crystallization of the hard segment of the UA series was completely inhibited as the grafted cholesterol content increased, which were supported by dynamic mechanical analysis (DMA) results for the storage modulus. As the cholesterol content increased, the glass transition temperature (T g ) of the UA series increased and remained the same for the UB series. The tensile strength in the UA series sharply increased with the cholesterol content, unlike that of the UB series. The strain at break in the UA series remained the same as the cholesterol content increased, whereas that of the UB series decreased significantly. As the cholesterol content increased, the shape recovery of the UA series remained above 90% at 45 °C and sharply increased at 0 °C. Finally, the UA series containing grafted cholesterol demonstrated excellent low temperature flexibilities compared to the UB series and unmodified PU.