Tea is one of the most widely consumed beverages in the world and may be associated with reduced heart disease rates. Theaflavins, which are formed in the production of black tea, have been suggested being responsible for the blood-cholesterol-lowering (BCL) effects of tea. We hypothesized that the effect of theaflavins on BCL could be through interference in the formation of dietary mixed micelles, which could result in reduced intestinal cholesterol absorption. Micelles were produced by mixing oleic acid, bile acids, lyso-phosphatidylcholine, and cholesterol. Theaflavin-treated micelles/particles were analyzed using electron microscopy (cryo-TEM), high-performance liquid chromatography (HPLC) analysis, and light-scattering particle size measurements. A dose-dependent inhibitory effect of theaflavins on the incorporation of 14C-labeled cholesterol into micelles and a theaflavin-dependent increase in particle size was found. These particles consisted of insoluble large multilamellar vesicles with onion-like structures. Ultracentrifugation and HPLC analysis revealed that the pellets contained mainly theaflavin-3-gallate, while the remaining theaflavins were found to be present in the supernatant. Using purified theaflavin subtypes confirmed that mainly theaflavin-3-gallate is responsible for multilamellar vesicle formation. These results show that theaflavins can play a role in decreased intestinal cholesterol absorption via inhibition of micelle formation.