4t1 mouse mammary tumor cell line

The IGROV-1 human ovarian cancer cell line (kind gift from Dr. J. Benard, Institute Gustave Roussy, Villejuif, France), 4T1 mouse mammary tumor cell line (American Type Culture Collection, ATCC, Manassas, VA, USA), EL-4 and YAC-1 mouse lymphoma tumor cell lines (ATCC, Manassas, VA, USA) were cultured in RPMI 1640 medium (Thermo Fisher Scientific Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific Inc., Waltham, MA, USA) and 2 mM glutamine (Thermo Fisher Scientific Inc., Waltham, MA, USA). The B16 mouse melanoma cell line (ATCC, Manassas, VA, USA) and RAW264.7 macrophage-like cell line (ATCC, Manassas, VA, USA) were maintained in DMEM (Thermo Fisher Scientific Inc., Waltham, MA, USA) supplemented with 10% FBS (Thermo Fisher Scientific Inc., Waltham, MA, USA) and 2 mM glutamine (Thermo Fisher Scientific Inc., Waltham, MA, USA). All the cell lines were routinely maintained at 37 °C in a 5% CO₂ atmosphere. The cell lines were authenticated by the Genomic Facility at Fondazione IRCCS Istituto Nazionale dei Tumori (Milan, Italy) and regularly tested for Mycoplasma using a specific kit (MycoAlert^TM Mycoplasma Detection Kit, Lonza Group Ltd., Basel, Switzerland).

Mouse mammary tumor cell line Mvt-1, which was derived from a primary mammary tumor of a MMTV-VEGF/Myc bitransgenic mouse [36 (link)], was obtained as a gift from Dr. Danny Welch (KUMC) after written permission from Dr. Kent Hunter (NIH). The 4T1 mouse mammary tumor cell line, a TNBC type with highly metastatic cells derived from a spontaneously arising BALB/c mammary tumor [25 (link)], was obtained from American Type Culture Collection (ATCC, Rockville, MD, Cat# ATCC^® CRL-2539^™). Cells were cultured in a monolayer in DMEM supplemented with 10% FBS, 2 mM glutamine, 100 unit/ml penicillin, and 100 unit/ml streptomycin in a 37°Cincubator in the presence of 5% CO₂. Cells were used between four to six passes for any experiment and were checked every two months for mycoplasma contamination.