MDA-MB-231 (human triple-negative breast cancer) and MCF10A (human nonmalignant breast epithelial) cell lines were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA). MDA-MB-231 were grown in Dulbecco’s Modified Eagle Medium (DMEM) with GlutamaxTM supplemented with 10% heat inactivated fetal bovine serum (FBS) and antibiotics (100 U/mL streptomycin, 100 μg/mL penicillin G). MCF10A cells were cultured in DMEM/F12 medium supplemented with 5% horse serum, 20 ng/mL EGF, 0.5 mg/mL hydrocortisone, 100 ng/mL cholera toxin, 10 μg/mL insulin, and antibiotics.
Human macrophages were derived from monocytes extracted from fresh buffy coats of healthy donors (obtained from Azienda Ospedaliera Padova, Padova, Italy) by centrifugation over a Ficoll– Hypaque step gradient and subsequent Percoll gradient (Sigma-Aldrich, Munich, Germany).
All cell culture medium and supplements were purchased from Life Technologies or Sigma-Aldrich (Munich, Germany), while sterile plasticware was purchased from Falcon® (Corning, New York, NY, USA).
Human macrophages were derived from monocytes extracted from fresh buffy coats of healthy donors (obtained from Azienda Ospedaliera Padova, Padova, Italy) by centrifugation over a Ficoll
All cell culture medium and supplements were purchased from Life Technologies or Sigma-Aldrich (Munich, Germany), while sterile plasticware was purchased from Falcon® (Corning, New York, NY, USA).