Culturing Neuroblastoma and Monocyte Cell Lines

The human malignant neuroblastoma SK-N-BE(2), SH-SY5Y cell lines, and human monocytic cell line THP-1 were obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Science (CBTCCCAS), Shanghai, China. The cell lines were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) (Hyclone, Logan, USA) supplemented with 10% fetal bovine serum (FBS) (Hyclone, Logan, USA). Cells were cultured at 37°C with 5% CO₂ in a humidified atmosphere. Recombinant human TNF-α (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in 0.1% bovine serum albumin (BSA) in PBS and stock solution (10 μg/ml) was stored at −20°C. Recombinant human SDF-1α (Life Technologies, Carlsbad, CA, USA) was prepared in 0.1% BSA in PBS and stock solution (100 μg/ml) was stored at 4°C. PMA (phorbol myristate acetate), a THP-1 cells inducer (Sigma-Aldrich, St. Louis, MO, USA), was prepared in PBS (320 nM) and kept at 4°C until used. PDTC (pyrrolidinedithiocarbamic acid ammonium salt), a NF-κB(p65)inhibitor [13 (link)] (Sigma-Aldrich, St. Louis, MO, USA) was prepared in PBS (50μM) and kept at 4°C until used.