A procedure based on freezing and thawing was developed to induce the rupture of adsorbed lipid vesicles on solid surfaces into supported lipid bilayers (SLBs). The SLB assembly exploits the phase transition of both lipids and water during freezing. It enables SLB formation independent of the type of substrates and lipids as long as the vesicles spontaneously adsorb onto the surface. The created SLB is a single bilayer, and has a diffusion coefficient of (0.6–4) × 10−8 cm2 s−1 on TiO2, which is in the same range as the SLBs formed by conventional techniques. The presented approach has the advantages of both the Langmuir–Blodgett method (the versatility in the selection of lipids and substrates) and vesicle fusion (self-assembly) at the same time.