Ecotoxicological evaluation of chemicals for European REACH regulation: a manual for growth and maintenance of the established fish cell line RTG-2 (Rainbow Trout Gonad)
In the frame of European Regulation REACH (CE n. 1907/2006), all chemical substances with a volume of production higher than 1 t per year, must be tested for their ecotoxicity. In addition to tests with algae and crustaceans, a large number of toxicity tests with fish has to be conducted.
Due to the good correlation found between the in vitro data and the in vivo fish data, the use of established fish cell lines can represent an alternative tool to acute fish bioassay for toxicity screening of chemicals.
In order to reduce and to supplement in vivo animal tests, a project of the Environmental Metrology Service of ISPRA, entitled “In vitro cytotoxicity assays for ecotoxicological assessment of chemicals: use of RTG-2 (Rainbow Trout Gonad) cell line”, is presented. The main goal of this project is the development and dissemination of alternative in vitro methods to the laboratories of Regional Environmental Protection Agencies of Italy (ARPA/APPA), involved in REACH application. The utilization of RTG-2 cell line has many practical advantages: can be incubated at room temperature and ambient atmosphere, is a rapid and inexpensive tool for routine basal cytotoxicity testing. Growth and maintenance conditions (basic laboratory equipment, culture medium, supplements, harvesting anchorage-dependent cells, growth curve, determination of cell concentration) of the cell line RTG-2 are presented (phase I of the project).