Investigation of culture conditions for endothelial cells retained using microbubbles and a controlled interferential acoustic field

In regenerative medicine, artificial organ development requires vascular networks, which remain difficult to reproduce with existing fabrication methods. We previously proposed bubble-surrounded cells (BSCs), in which lipid microbubbles attach to endothelial cells and are manipulated by ultrasound. In this study, we examined cell retention and subsequent in situ culturing under interferential acoustic fields. Endothelial cells conjugated with lipid bubbles were retained on the upper surface of a flow path and cultured for several days under different acoustic conditions. Phase sweeping reduced cell damage and promoted stable culture, whereas continuous exposure without sweeping caused cell loss. These findings demonstrate that optimized acoustic fields enable stable retention and multiday in situ culture, supporting applications in the construction of artificial blood vessels.