Primary epithelial BC and FAD cultures were prepared from biopsies obtained from patients at Clinica Mediterranea S.p.A, as previously reported [13 (link)]. Our study included two biological replicates and three technical replicates for each patient-derived EV preparation, whereas there were three technical replicates for each model-derived EV preparation.
Cell cultures were grown in Dulbecco’s modified Eagle’s medium (DMEM)/Nutrient F12-Ham (DMEM-F12) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, Milan, Italy). Continuous cell lines were purchased from ATCC (LG Standards, Milan, Italy) and grown in the following media: Roswell Park Memorial Institute medium (RPMI) for human breast BT549 and MDA-MB-231 cells; DMEM/F12 supplemented with EGF (20 ng/mL), hydrocortisone (0.5 μg/mL), cholera toxin (100 ng/mL), insulin (10 μg/mL), and horse serum (5%) for human breast MCF–10A cells. All cells were grown at 37 °C in 95% air and 5% CO2.
Cell cultures were grown in Dulbecco’s modified Eagle’s medium (DMEM)/Nutrient F12-Ham (DMEM-F12) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, Milan, Italy). Continuous cell lines were purchased from ATCC (LG Standards, Milan, Italy) and grown in the following media: Roswell Park Memorial Institute medium (RPMI) for human breast BT549 and MDA-MB-231 cells; DMEM/F12 supplemented with EGF (20 ng/mL), hydrocortisone (0.5 μg/mL), cholera toxin (100 ng/mL), insulin (10 μg/mL), and horse serum (5%) for human breast MCF–10A cells. All cells were grown at 37 °C in 95% air and 5% CO2.