Success in fibroblast cell cultures from long-beaked common dolphin (Delphinus capensis) and bryde's whale (Balaenoptera edeni) skin biopsies of the Gulf of California (Mexico): potential application for ecotoxicological studies

The aim of this study was to develop fibroblast cell cultures from skin biopsies of free-ranging cetaceans from the Gulf of California (Sea of Cortez-Mexico): long-beaked common dolphins (Delphinus capensis) and Bryde's whales (Balaenoptera edeni), as methodological tool in ecotoxicological studies. The biopsy dart, a regular aluminium crossbow bolt with a modified stainless steel collecting tip, was used in different ways depending on the species. The tissue was kept in tissue culture medium at ambient temperature and processed in 72h. The main result was the success for the first time in fibroblast cultures of two long-beaked common dolphin and two Bryde's whale specimens. The growth of first fibroblasts were observed after 10 days both in dolphins and in whales. The difference was that fibroblast cultures reached 90% confluence in 50 ml Falcon flasks in 15 days time for long-beaked common dolphins while fibroblasts from Bryde's whales grew slowly and do not reached confluence in 50 ml flasks. The dolphin fibroblasts were trypsinized, washed and placed in 250 and 550 ml flasks, after two and three trypsinizations, respectively. The cultures thus obtained can be used for many purposes, including genetic, biochemical and toxicological studies. In particular, fibroblasts can be used to test the susceptibility of these cetaceans to different environmental contaminants such as organochlorine compounds (OCs) and polybrominated diphenyl ethers (PBDEs). Fibroblast cell cultures of Mexican cetaceans, treated with different mixtures of OCs, PBDEs and PAHs, will be analysed by immunofluorescence and western blotting techniques for a qualitative and quantitative evaluation of target proteins such as CYP1A1-1A2 and CYP2B4.