The adherent fraction of isolated PBMCs was cultured for 4 to 5 days on tissue culture plastic with medium replacement every 1 to 2 days before experiments with silicones. Liquid silicone (SYLGARD 184, Dow Corning) was spread on the bottom of wells in a 96-well plate. Following an incubation time of 90 min at 70°C of the liquid silicone, cells (100,000 cells per well for 96-well plates) were seeded on top of the liquid silicone and allowed to adhere for 45 to 60 min at 37°C and 5% CO2. Adherent cells were covered with a previously prepared collagen I master mix supplemented with cell culture medium to obtain a final concentration of 1.6 mg/ml. Polymerization of the collagen gel and equilibration of gas conditions within the collagen matrix took place for 20 to 30 min at 37°C and 5% CO2 to obtain a final pH of 7.4 to 7.5. The pH values were determined using indicator paper strips. Cells attached to glass and silicone and covered with fibrillar collagen I matrices were maintained for 7 days with medium replacement every 1 to 2 days. For treating cells with inhibitors, dasatinib (11498, Cayman Chemical) and PF-573228 (14924, Cayman Chemical) were used at final concentrations of 0.1 and 5 μM, respectively, for 7 days.

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