BioProject

LX2 cells treated with or without BTC and with or without oleic acid or DHA Overall design: LX2 cells were obtained from SL Friedman (Mount Sani Medical School) (46). More...

LX2 cells treated with or without BTC and with or without oleic acid or DHA Overall design: LX2 cells were obtained from SL Friedman (Mount Sani Medical School) (46). LX2 cells are activated human hepatic stellate cells; they were maintained in DMEM with 10% FCS containing penicillin and streptomycin. Cells were plated onto 100 mm plastic petri dishes ∼100,000 cells/plate and treated with fatty acids (at 50 μM) in endotoxin-free BSA (at 20 μM) during the growth phase. Fatty acids [oleic acid (18:1 ω9) and DHA (22:6, ω3); Nu-Chek Prep] were used at 50 μM for 2- 48-hour cycle treatments. After this pre-treatment, cells were trypsinized, washed in PBS, counted and plated at 3000 cells/well in a 96-well plate. Cells were fed DMEM with 1 % FCS without or with betacellulin (human recombinant betacellulin (BTC), R&D Systems) for 72 hrs; the concentration ranged from 1.25 to 25 ng/ml. At the completion of BTC treatment, media was removed, washed with PBS and DNA/well was quantified using the CyQuant cell proliferation assay (Thermofisher); DNA fluoresces was quantified [excitation at 485 nm & emission at 530 nm. This experiment was repeated 3 times. Less...