Thermo lab systems plate reader

Theca cells were seeded into 96‐well plates and cultured overnight with McCoy's medium containing 0.1% BSA. After attachment, cells were pretreated with IL‐18 (300 pg/mL) for 1 hour followed by LH (150 pg/mL) for 24 hours. After the treatment periods, cell viability was determined by MTT assay.³¹ For cell proliferation assay, TCs were treated with 10 μmol/L bromodeoxyuridine/5‐bromo‐2’‐deoxyuridine (BrdU) in the presence or absence of IL‐18 (0, 10, 30, 100, 300, 500 and 1000 pg/mL), cell proliferation was measured using an enzyme‐linked immunosorbent assay after 48 hours (Cell Proliferation ELISA, BrdU [colorimetric], Roche Applied Science), according to the manufacturer's recommendations. The absorbance was measured at 450 nm with a Thermo Lab Systems plate reader (Thermo Fischer Scientific) and Ascent Software for Multiskan equipment. Cell proliferation was normalized to the control condition of each culture. The results are expressed as mean ± standard error of the mean (SEM) of four independent experiments with three replicates per condition.

After 48-h treatment with 10 µM bromodeoxyuridine/5-bromo-2’-deoxyuridine (BrdU) in the presence or absence of BPS (10 nM, 100 nM, 1 µM, 10 µM or 50 µM) and removal of the supernatant, cell proliferation was measured using an enzyme-linked immunosorbent assay (Cell Proliferation ELISA, BrdU [colorimetric], Roche Applied Science, Germany), according to the manufacturer’s recommendations. The absorbance was measured at 450 nm with a Thermo LabSystems plate reader (Thermo Fischer Scientific) and Ascent Software for Multiskan equipment. Cell proliferation was normalised to the control condition of each culture. The results are expressed as mean ± standard error of the mean (SEM) of five independent experiments with four replicates per condition.