Cyclodextrins are widely used excipients to increase water solubility and bioavailability of lipophilic drugs. Our research group previously showed that cyclodextrins are able to enter Caco-2 intestinal epithelial cells by endocytosis, however, the cellular internalization and intracellular effects in the case of different cells have not been investigated yet. In addition, 2-hydroxypropyl-beta-cyclodextrin (HPBCD) is a potential molecule for the treatment of Niemann Pick syndrome type C due to its cholesterol complexing properties. The aim of our work is to investigate and compare the cellular internalization of HPBCD and its effect on lysosomes on Caco-2 intestinal epithelial, HeLa cervical epithelial and hCMEC/D3 brain endothelial cells.