Stimulating NIH3T3 Cells with Growth Factors

NIH3T3 cells were cultured in Dulbecco's modified Eagle's medium (DMEM, from Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Scientific, Waltham, MA, USA) and maintained in humidified atmosphere of 5% CO₂ at 37^°C. For cell treatments, recombinant transforming growth factor-β (TGF-β) (5 ng/mL), tumor necrosis factor-α (TNF-α) (5 ng/mL), vascular endothelial growth factor (VEGF)-165 (20 ng/mL), platelet-derived growth factor (PDGF) (10 ng/mL) or interleukin-6 (IL-6) (10 ng/mL) (all purchased from R&D Systems, Minneapolis, MN, USA) were added to the medium for 24 hrs. The macrophage cell line RAW264.7 was cultured in DMEM containing 10% FBS as described [48 (link)].