hOMC monolayer culture
PA5 hOMCs were expanded in T-flasks, freshly coated with poly-L-lysine
(PLL, Sigma, UK, 100 μg/mL), using as complete medium DMEM/F12 +
GlutamaxTM (Gibco, Life Technologies, UK) supplemented
with 10% Fetal Bovine Serum (FBS) (Sigma, Germany) and
4-Hydroxitamoxifen (4-OHT, 1:10,000). Cells were maintained in a
standard incubator (37⁰C, 5% CO2 in air). A seeding
density of 6000 cells/cm2 was used and feeding was
performed every two days. Cells were passaged when 60-80% confluency
was reached, through incubation with TryPLE (Gibco, UK) for 5 minutes.
The dissociation reagent was quenched with complete medium and the cell
suspension was centrifuges for 5 minutes at 400 x g.