Dmem f12 phenolredfree

The 12-Z cell line was a generous gift of Dr. Anna Starzinski-Powitz (Goethe-Universität Frankfurt) [38 (link)]. Human epithelial endometriotic cell line 12-Z was cultivated in DMEM-F12 phenolredfree (Gibco, Thermo Fisher Scientific, Vienna, Austria) with 10% (v/v) heat inactivated fetal calf serum (FCS, SigmaAldrich, Vienna, Austria) and 1% (v/v) penicillin and streptomycin (Sigma-Aldrich, Vienna, Austria). Cultivation was done in humidified incubators at 37 °C and 5% CO₂. The 12-Z cell line was grown in T75 polystyrene cell culture flasks with cell growth surface for adherent cells (Sarstedt, Wiener Neudorf, Austria). Cells were sub-cultured every 3–4 days at a 1:3 ratio. Cells were cultivated until they reached a concentration of 80% confluency. Cell counting was performed using a MOXI Z mini automated cell counter (ORFLO Technologies, Ketchum, ID, USA) using Moxi Z Type M cassettes (ORFLO Technologies, Ketchum, ID, USA). Cells were routinely checked for mycoplasma contamination using MycoAlert™ mycoplasma detection kit (Szabo-Scanidc, Vienna, Austria). Cells were seeded on a 6-well plate with cell growth surface for adherent cells (Sarstedt, Wiener Neudorf, Austria) at a density of 300.000 cells/well in 3 mL growth medium. Inflammatory stimulation was applied for 48 h at a concentration of 10 ng mL⁻¹ with either TNFα or IL1β (both Sigma-Aldrich).

Fluo-4 (F14200), Fluo-4 AM (F14201), Dextran Texas Red, 3,000, 10,000, and 70,000 MW (D3329, D1828, and D1830, respectively) were purchased from Thermo Fisher Scientific. CT04 and CN03 were the products of Cytoskeleton Inc. Cytochalasin D (C2618), cytosine arabinofuranoside (Ara-C), poly-D-lysine, and paraformaldehyde were purchased from Millipore Sigma. NGF was purified from mouse salivary glands. Goat anti-mouse Alexa 488, 10% goat serum, and laminin (1 μg/mL) were products of Life Technologies. Mouse Tuj1 primary antibody was the product of Covance. DMEM high glucose (11,965,092), DMEM/F-12 (11,320,033), DMEM/F-12 phenol red free (21,041,025), DMEM high glucose, calcium free (21,068,028), 100X penicillin/streptomycin (10,378,016), and FBS (16,000,044) were brought from Gibco.