The functionalized triblock polymer for the preparation of self-assembly biomaterials, Chol-(CL) n -Chol, was synthesized through ring-opening polymerization of ecaprolactone (e-CL) following by chain-extension reaction. The chemical structure of functionalized triblock polymer was confirmed by Fourier transform infrared and 1 H-NMR. The molecular weight of the functionalized polymer increased with decreasing feed ratio of the initiator cholesterol (Chol) to the monomer e-CL. Liquid crystalline properties and the self-assembly behavior of the functionalized polymer were validated by Wide angle X-ray diffraction, differential scanning calorimetry, and polarizing light microscope. The results showed that Chol-(CL) n -Chol ( n 40) exhibited liquid crystallinity in particular temperature ranges because of the incorporation of the cholesteryl moiety. In addition, the functionalized triblock polymer can be homogeneously blended with high molecular weight poly(e-caprolactone) (PCL), which offers the possibility to modify the properties of biomaterials based on PCL. Mixing a small amount of a Chol-(CL) n -Chol functionalized polymer with PCL, or coating a thin layer of the functionalized polymer on the surface of a PCL-based implant may modify cell response.