4.10. Murine Primary PDAC Cell Culture, Cytokine and Inhibitor Treatment

The mouse primary pancreatic cancer cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, nonessential amino acids, 100 units/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a 5% CO₂ incubator. Primary mouse PDAC cells were maintained for <6 passages and histopathologically characterized through SCID mice xenograft studies before performing microarray expression profile analyses. Tumor spheroids were created using Perfecta3D^® Hanging Drop Plates (Sigma Aldrich, St. Louis, MO, USA). Spheroids of cells (2 × 10³ cells) were prepared as described above. PDAC cells were treated with 20 mM lithium chloride (LiCl) (Sigma-Aldrich 203637) for 18 h before Western blot analysis.