Breast Cancer Cell Line Maintenance and Stimulation

The human breast cancer cell lines MDA-MB-231 (ATCC® HTB-26™), BT549 (ATCC® HTB122™), HCC1937 (ATCC® CRL 2336™), MDA-MB-468 (ATCC ® HTB-132™), (American Type Culture Collection, Manassas, VA), SUM149, and SUM159 (Asterand Bioscience, Detroit, MI now acquired by BioreclamationIVT, Westbury, NY) were authenticated using a panel of microsatellite markers. Cell lines were maintained at 37 °C in a humidified atmosphere of 5% CO₂ as previously described in Turdo et al. [3 (link)]. For stimulation experiments, MDA-MB-231 cells were starved in serum-free medium for 24 h and then treated for 48 h with a pool of 5 WHFs at a final concentration of 5% as described [23 (link)] or with PDGF-BB, Mib1b, MCP1, IP10, Il1ra, Il1b, G-CSF, Il8, Il6, EGF, FGF, Heregulin, PDGF-AA, PDGF-AB (PeproTech, Rocky Hill, NJ) at 50 ng/mL. Cells were treated in indicated experiments with cycloheximide (1 μM) or UO126 (2 μM), both of which were dissolved in DMSO (maximum concentration 0.1%) (Sigma-Aldrich).