Pennicilin streptomycin

Hyclone DMEM-F/12 1:1 (Thermo Scientific #SH30023.01) supplemented with:
10 mM HEPES (Sigma-Aldrich; #H3885),
10 μg/mL Gentamycin (Euroclone #ECM0011B; 10 mg/mL),
2 mM L- Glutamine (Euroclone ECB3000D-20; 200 mM),
1% Pennicilin/streptomycin (Euroclone #ECB3001D, 100×),
2.5 μg/mL Amphotericin B (Euroclone #ECM00 09D; 250 μg/mL),
5 mM Nicotinammide (Sigma-Aldrich, #N0636, 250 mM).
1.25 mM N-acetylcysteine (Sigma-Aldrich, # A9165, 125 mM).
1× B27 supplement (Gibco, #17504–44, 50×).
250 ng/mL R-spondin 3 (R&D, #3500-RS/CF, 25 μg/mL)*.
5 nM Heregulin (Peprotech, #100–03, 7.14 μM)*.
5 ng/mL KGF (Peprotech, #100–19, 10 μg/mL)*.
20 ng/mL FGF10 (Peprotech, #100–26, 25 μg/mL)*.
5 ng/mL EGF (Peprotech, #AF-100-15, 5 μg/mL)*.
100 ng/mL Noggin (Peprotech, #120-10C, 20 μg/mL)*.
500 nM A83–01 (Tocris, #2939, 500 μM)*,**.
5 μM Y-27632 (Abmole,#M1817, 5 mM)*.
500 nM SB202190 (Sigma-Aldrich, #S7067, 500 μM)*.
* Add fresh the day of use.
** dissolve in DMSO.

Briefly, a TNBC tissue was digested by Collagenase I, then cultured in DMEM‐F/12 (Gibco) supplemented with 10 mm HEPES (Sigma‐Aldrich), 10 µg mL⁻¹ Gentamycin (Euroclone), 2 mm L‐Glutamine (Euroclone), 1% Pennicilin/streptomycin (Euroclone), 2.5 µg mL⁻¹ Amphotericin B (Euroclone), 5 mm Nicotinamide (Sigma), 1.25 mm N‐acetylcysteine (Sigma), 1 × B27 supplement (Gibco), 250 ng mL⁻¹ R‐spondin 3 (R&D), 5 nm Heregulin (Peprotech), 5 ng mL⁻¹ KGF (Peprotech), 20 ng mL⁻¹ FGF10 (Peprotech), 5 ng mL⁻¹ EGF (Peprotech), 100 ng mL⁻¹ Noggin (Peprotech), 500 nm A83–01 (Tocris), 5 µm Y‐27632 (Abmole), 500 nm SB202190 (Sigma). The TNBC organoids were treated with CRT0066101 (1 µm) and/or OGX‐011 (300 nm) for cell growth assay.