Influence of type and quantity of serum on the effects of synthetic estrogen on Channel Catfish Ovary (CCO) cells

Za ispitivanje učinaka endokrinih modulatora prisutnih u okolišu sve više se rabe in vitro testovi. Pri ispitivanju in vitro citotoksičnosti, stanice se uzgajaju u mediju kojem je dodan serum životinjskog podrijetla. Vezanje na proteine seruma može značajno utjecati na dostupnost ispitivane kemikalije, a samim time i na odnos koncentracija-učinak. Stoga je cilj ovog rada bio ispitati utječe li vrsta i volumni udio seruma na djelovanje dietilstilbestrola (DES) i 17α- etinilestradiola (EE2) na CCO stanice. Stanice su tretirane DES-om i EE2 u koncentracijama od 0,1 do 10 µg/mL, u mediju za uzgoj s 2,5; 5 i 10 % (v/v) običnog telećeg seruma (FBS) ili seruma obrađenog aktivnim ugljenom (Cs-FBS). Nakon 72 sata od tretmana, vijabilnost CCO stanica određena je WST-1 metodom. Na temelju dobivenih rezultata i izračunatih EC50 vrijednosti pokazano je da učinak DES-a i EE2 na CCO stanice ovisi o vrsti i volumnom udjelu seruma u mediju za uzgoj.The in vitro tests are frequently used to determine the impact of endocrine modulators present in the enviroment. During the in vitro testing of citotoxicity, the cells are grown in the media with added serum of animal origin. Binding to serum proteins could have an impact on the availability of tested chemicals and therefore on the concentration-effect ratio. The aim of this study was to test if the type and quantity of serum have influence on the diethylstibesterol (DES) and 17α-ethynylestradiol treatment of CCO cells. The cells have been treated with DES and EE2 concentrated between 0,1 and 10 μg/mL, in a medium with 2,5; 5 or 10% (v/v) fetal bovine serum (FBS) or serum stripped with charcoal (Cs-FBS). After 72 hours of tretment, the viability of CCO cells has been determined by WST-1 method. The obtained results and calculated EC50 values showed that the impact of DES and EE2 on CCO cells depend on the type and quantity of serum aded in the cultivation medium