This chapter examines different methodology for maintaining elastin-producing cells in culture, and then considers factors that modulate elastin production. While studies of this kind have provided important insights into regulatory pathways in cultured cells, it should be stressed that the elastin phenotype is extremely unstable in vitro and that conclusions from manipulation of cultured cells may not be relevant for cells in situ until direct comparison of both conditions is possible. Nevertheless, cell culture techniques remain one of the most useful tools for studying elastin metabolism. Numerous cell types synthesize elastin in vitro, including aortic smooth muscle cells, dermal fibroblasts, fibroblasts from ligamentum nuchae, ear chondroblasts, lung mesothelial cells, bovine corneal endothelial cells, and vascular endothelial cells. All of these cell types express the elastin phenotype when maintained under appropriate culture conditions although it should be emphasized that how one assays for elastin is an important parameter in assessing levels of elastin synthesis.