Nanomedicines are believed to be capable of tumor-selective drug delivery, but their clinical application has not progressed. In this study, we aimed to construct a microfluidic tumor model that reflects the structure and characteristics of human tumor tissue and apply it to the evaluation of nanomedicines. Vascular endothelial cells, tumor cells, and macrophages were incorporated into the device, and the permeability and cellular uptake of nanoparticles were evaluated. Microfluidic devices suitable for microscopic observation were fabricated, and the effects of various conditions on the dynamics of nanoparticles were clarified.