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.
Epithelial cell culture protocol
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.
Corresponding Organization : Istituto Neurologico Mediterraneo
Other organizations : Institute for Experimental Endocrinology and Oncology, National Research Council, University of Ottawa
Variable analysis
- Primary epithelial BC and FAD cultures prepared from biopsies obtained from patients at Clinica Mediterranea S.p.A
- Continuous cell lines (BT549, MDA-MB-231, MCF-10A) purchased from ATCC
- Extracellular vesicle (EV) preparations
- Cell cultures grown in DMEM/Nutrient F12-Ham (DMEM-F12) supplemented with 10% fetal bovine serum (FBS)
- Continuous cell lines grown in the following media: RPMI for BT549 and MDA-MB-231, DMEM/F12 supplemented with EGF, hydrocortisone, cholera toxin, insulin, and horse serum for MCF-10A
- All cells grown at 37 °C in 95% air and 5% CO2
- Two biological replicates and three technical replicates for each patient-derived EV preparation
- Three technical replicates for each model-derived EV preparation
Annotations
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