Anthropogenic activities have increasingly contaminated aquatic ecosystems worldwide, requiring the development of adequate methods to assess the effects of environmental pollution on aquatic biota. Currently, ecotoxicological research on fish is largely based on in vivo studies, many times using post-mortem fish samples bought in fish markets or obtained through capture-and-release programs. However, such samples provide a narrow window to the cellular and molecular processes that occur to fish upon exposure to pollutants and other toxicants or pathogens. In thi sense, in vitro cell culture systems have been increasingly proven a valuable tool in several research fields, from molecular biology studies to conservation efforts. To date, however, cell cultures obtained from bony fish have been the most studied and with the best-described protocols and models. Elasmobranchs, comprising shark and rays, play important trophic and environmental roles, employed as chemical contamination environmental sentinels, suffering the effects of such contamination due to bioaccumulation and biomagnification processes. For these reasons, the development of new experimental tools to study elasmobranch cellular and molecular responses to environmental stimuli in controlled conditions is highly desirable. However, only some research groups have attempted to develop elasmobranch cell culture protocols to be used in an ecotoxicological context. In this sense, this review discusses the current elasmobranch cell culture scenario, its importance and potential applications in ecotoxicology assessments and conservation actions.