Bio myc

TEVA was performed once the volume of the PDX (#7157) had reached ~500 mm³. The PDXs were excised aseptically from the mice and cut into 2 × 2 × 2 mm³ tissue explants. The 2 × 2 × 2 mm³ explants were then incubated with the different drugs for 24 h in 48-well tissue culture plates at 37 °C, 95% relative humidity and 5% CO₂, under sterile conditions with suitable controls. The 2 × 2 × 2 mm³ explants incubated only in culture media without any drug served as control. DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 20% FBS (Gibco), 1 mM sodium pyruvate (Biological Industries, Haemek, Israel), 2 mM l-glutamine (Biological Industries), 1% penicillin–streptomycin–amphotericin (Biological Industries), 0.1 mM MEM non-essential amino acids (Biological Industries), 10 mM HEPES (Biological Industries), 1% BIO-MYC (Biological Industries), and 50 µg/mL gentamycin (Gibco) was used as the culture medium. The therapeutic agents for this part of the study were BYL719 (2 µM), GDC0032 (500 nM), and AEW541 (1 µM). KI67, TUNEL, and pMAPK staining were measured and the TEVA score was calculated using the following formula: (0.3 × KI67 staining) + (0.3 × TUNEL staining) + (0.3 × MAPK staining).

When the PDXs (preferably first generation PDX) reached ~500 mm³, they were excised from the mice. Tumors were then cut into 2 × 2 × 2 mm³ tissue explants and cultured on gelatine sponges in 12-well tissue culture plates for specific time points as indicated in the text. The DMEM culture media (Gibco) containing 20% FBS (Gibco), 1 mM sodium pyruvate (Biological Industries), 2 mM l-glutamine (Biological Industries), 1% penicillin/streptomycin/amphotericin (Biological Industries), 0.1 mM MEM non-essential amino acids (Biological Industries), 10 mM HEPES (Biological Industries), 1% BIO-MYC (Biological Industries) and 50 μg ml⁻¹ gentamicin (Gibco). For drug treatment, the 2 × 2 × 2 mm³ explants were treated with 3 μM BAY-876 for indicated time in 37 °C, 5% CO₂.

When the PDXs (preferably first generation PDX) reached ~500 mm³, they were subjected to Tumor ex vivo Analysis (TEVA). After excising out the PDXs aseptically from mice they were cut into 2 × 2 × 2 mm³ tissue explants and cultured in 48 well tissue culture plates for specific time point as indicated in the text. The DMEM culture media (Gibco) contained 20% FBS (Gibco), 1 mM sodium pyruvate (Biological Industries), 2 mM L-glutamine (Biological Industries), 1% penicillin/streptomycin/amphotericin (Biological Industries), 0.1 mM MEM non-essential amino acids (Biological Industries), 10 mM HEPES (Biological Industries), 1% BIO-MYC (Biological Industries) and 50 ug/ml gentamycin (Gibco). For drug treatment, the 2 × 2 × 2 mm³ explants were treated with different therapeutic drugs for 24 h in 48 well tissue culture plates in 37°C, 5% CO₂.