Localized drug delivery with ultrasound-induced hyperthermia can enhance the therapeutic index of chemotherapeutic drugs by improving efficacy and reducing systemic toxicity. A novel in vitro method for the activation of drug-loaded thermosensitive liposomes is described. In particular, a dual-compartment, acoustically transparent container is used in which thermosensitive liposomes suspended in cell culture medium are immersed in a thermally absorptive medium, glycerol. Hyperthermia is induced with ultrasound in the glycerol, which in turn heats the culture medium by thermal conduction. The method approximately mimics the in vivo scenario of thermosensitive liposomes collected in the interstitial spaces of tumors, where ultrasound induces hyperthermia in the tumor tissue, which in turn heats the thermosensitive liposomes by conduction and induces release of the encapsulated drug. The acoustic conditions for the desired hyperthermia are derived theoretically and validated experimentally. Eighty percent release of doxorubicin from thermosensitive liposomes is achieved.