Based on their ability to exchange lipids between vesicles, cyclodextrins (CD) have been identified that can catalyze the formation of artificial asymmetric vesicles (AsV). While modified cyclodextrins have been studied for their ability to assemble AsV in the past, unmodified cyclodextrins, such as alpha-cyclodextrin (alphaCD) and gamma-cyclodextrin (gammaCD) have not been actively examined for such applications. In this study, we tested alphaCD and gammaCD, to understand their interaction with lipids and to evaluate their applications in AsV assembly. While both cyclodextrins showed little ability to solubilize lipids from optical density measurements, both exhibited a significant extent of CD-mediated lipid exchange ability as assay by FRET (Fluorescence Resonance Emission Transfer). The data suggest that alphaCD and gammaCD have a significant ability to interact with lipid and have the potential for constructing AsV. In experiments using alphaCD and gammaCD to prepare asymmetric vesicles, both cyclodextrins showed the ability to induce donor contamination of AsV, which are isolated after exchange by pelleting upon centrifugation, as judged a small amount of donor lipids pelleting in the absence of acceptor (alphaCD > 7mM, gammaCD > 80mM). Using lower dosage, gammaCD was able to prepare AsV with reasonable efficiency while avoiding contamination with donor lipid, whereas alphaCD resulted in low exchange efficiency. Therefore, gammaCD has more potential to be used in assembly of AsV than alphaCD. After optimizing exchange conditions to get even more efficient exchange, preparation of AsV using gammaCD with various lipid compositions may be tested in the future.