Dulbecco s phosphate buffered saline dpbs

Testing was performed in an adherent glial cell line cultured from the brain of rainbow trout (Onchorhynchus mykiss) named RTbrain-W1, which has seen limited use in the literature (Fischer et al., 2011; (link)Liu et al., 2011; (link)Lončar et al., 2010; (link)Steinmoeller et al., 2009; Vo et al., 2015) (link). Cells were routinely grown in 75 cm 2 (T75) polystyrene tissue culture flasks (BioLite, Thermo Fisher Scientific) at room temperature (RT; 20 ± 2 °C) in L-15 basal medium (HyClone, GE Healthcare) supplemented with 15% fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific) and a 1% penicillin-streptomycin cocktail (P/S; HyClone, GE Healthcare). Routine passaging and seeding was performed using 0.25% v/v trypsin (HyClone, GE Healthcare) diluted in Dulbecco's phosphate-buffered saline (DPBS; from HyClone, GE Healthcare), and cells were used between their 5 th and 25 th passages.
Prior to exposing cells to genotoxicants or solvent only for controls, cells were removed from T75s by trypsinization and centrifuged (3,000 RPM for 5 min at 18 °C). The trypsin was removed, and cells were seeded on either a 96-well plate or a 6-well plate (BioLite, Fisher Scientific) for cytotoxicity tests or western blots, respectively, in L-15 with 15% FBS and 1% P/S for 24 h at RT. All genotoxicant exposures occurred in medium of L-15 with 10% FBS and 1% P/S.