Transwell is a device widely used for simulating mucosal and physiologic barrier systems, such as human reproductive tract mucosa, rectal mucosa, blood-brain barrier and blood-retina barrier in vitro. This device has been used to study he process of human immunodeficiency virus (HIV) entering into the body and to help us to illustrate , the mechanism of HIV infection. The system is also being used to facilitate the discovery of new vaccines and drugs for HIV. This review covers the current progresses of transwell applications in HIV-related studies.