This review will focus on the transport and intracellular formation of S-nitrosothiols in cell culture models. The major points made in this article are: (1) S-Nitrosothiols are actively metabolized by cells. (2) S-Nitrosothiols affect cells in ways distinctly different from those of nitric oxide and can act through mechanisms that do not involve the intermediacy of nitric oxide. (3) Some S-nitrosothiols (S-nitrosocysteine, S-nitrosohomocysteine) can be taken up into cells via amino acid transport system L, whereas others (S-nitrosoglutathione, S-nitroso-N-acetylpenicillamine) are not directly transported, but require the presence of cysteine and/or cystine before the nitroso functional group is transported. (4) Proteomic detection of intracellular S-nitrosothiols is currently possible only if cells are loaded with high levels of S-nitrosothiols, and methodological advances are required in order to examine the S-nitrosated proteome after exposure of cells to physiological levels of nitric oxide.