Cytotoxicity Assays with Fish Cells as an Alternative to the Acute Lethality Test with Fish

Abstract

In ecotoxicology, in vitro assays with fish cells are currently applied for mechanistic studies, bioanalytical purposes and toxicity screening. This paper discusses the potential of cytotoxicity assays with fish cells to reduce, refine or replace acute lethality tests using fish. Basal cytotoxicity data obtained with fish cell lines or fish primary cell cultures show a reasonable to good correlation with lethality data from acute toxicity tests, with the exception of compounds that exert a specific mode of toxic action. Basal cytotoxicity data from fish cell lines also correlate well with cytotoxicity data from mammalian cell lines. However, both the piscine and mammalian in vitro assays are clearly less sensitive than the fish test. Therefore, in vivo LC50 values (concentrations of the test compounds that are lethal to 50% of the fish in the experiment within 96 hours) currently cannot be predicted from in vitro values. This in vitro–in vivo difference in sensitivity appears to be true for both fish cell lines and mammalian cell lines. Given the good in vitro–in vivo correlation in toxicity ranking, together with the clear-cut difference in sensitivity, the role of cytotoxicity assays in a tiered alternative testing strategy could be in priority setting in relation to toxic hazard and in the toxicity classification of chemicals and environmental samples.

Proposal to improve vertebrate cell cultures to establish them as substitutes for the regulatory testing of chemicals and effluents using fish

“…Thus, DarT will likely play an important role in the development of alternatives to fish tests within the next years. An implementation at the regulatory level as substitute for a fish test has not yet been accomplished with fish or mammalian cultured cells (Kilemade and Quinn, 2003;Segner, 2004). However, numerous studies can be found that allow their potential and current limitations with regard to replacing fish tests to be evaluated.…”

Toxicological in vitro effects of heavy metals on gilthead seabream (Sparus aurata L.) head–kidney leucocytes

“…Therefore, more studies are needed to clarify the immunotoxicological effects and mechanisms of heavy metals in fish leucocytes. Furthermore, this kind of approximation could be helpful in fish immunotoxicology and should be explored, moreover with the knowledge that in vitro cytotoxicity assays with fish cell lines are very well correlated to acute lethality tests in vivo (Segner, 2004). Taking in consideration all the previous data, the aim of this work was to evaluate the immunotoxicological effects of four heavy metals (Cd, As, Pb and Hg) on the head-kidney leucocytes from gilthead seabream (Sparus aurata), the most important farmed fish species in the Mediterranean area.…”

Validation of an in vitro cytotoxicity test for four heavy metals using cell lines derived from a green sea turtle (Chelonia mydas)

“…The inherent economic and ethical constraints associated with in vivo tests using live animals for cytotoxicity analysis have led to the preferred utilization of in vitro biological test systems such as cell lines derived from aquatic animal species (Keddy et al 1995;Mátlová et al 1995;Huuskonen et al 1998;Segner 1998;Olabarrieta et al 2001;Repett et al 2001;Choi and Oris 2003;Ní Shúilleabháin et al 2004;Segner 2004;Wang et al 2004;Gűlden et al 2005 ) commonly present and bioavailable in the aquatic environment. This paper describes the employment of the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and CB (Coomassie Blue) assays for the examination of cytotoxic responses of these turtle cell lines following their exposure to the four selected metal salts.…”