Cell culture protocol for Breast Cancer cells

An isogenic panel of varying metastatic potential derived from the human breast cancer cell line MDA-MB-231 was used in this study. In addition to the parental MDA-MB-231 cells (P231), the panel consisted of CTC and LM cells previously obtained after orthotopic implantation of the parental cells in the fourth right mammary fat pad of female athymic nu/nu female mouse (NCI) as detailed in our previous publications [⁵⁹] [⁶⁰]. The three cell lines were cultured in RPMI-1640 media supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin and maintained at 37 °C and 5% CO₂ in a humidified incubator.