Thiols are a class of key compounds containing organic sulfur atoms, playing important physiological roles in living organisms. It can be ingested into organisms through various pathways, including active transport and passive diffusion mechanisms. Thiols participate in regulating various life activities such as antioxidant, detoxification, and protein synthesis in organisms. In recent years, thiol mediated uptake (TMU) has emerged as a cellular uptake mechanism, mainly through the formation of disulfide bonds between substrates and cell surface protein thiol groups, which then enter cells through various methods such as fusion, endocytosis, or direct transmembrane transport. Once inside the cell, the substrate can be released through dynamic exchange of disulfide bonds mediated by active small molecules such as GSH, thereby achieving delivery to some proteins, small molecule probes, and nanomaterials. Meanwhile, this uptake pathway can also be hindered by some thiol inhibitors, thereby hindering the uptake of thiol compounds. The latest research progress indicates that thiol mediated uptake has potential application prospects in the prevention and treatment of various diseases.