Mda mb 468

The TNBC cell lines MDA-MB-231 and MDA-MB-468 were purchased from American Type Culture Collection (ATCC, Manassas, VA). Cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin in a humidified atmosphere at 37°C with 5% CO₂ under atmospheric oxygen conditions (20%). Stable knockdown cells MDA-MB-468 were cultured in media supplemented with 2.5 μg/ml puromycin (Sigma-Aldrich, St. Louis, MO). For stable knockdown, cells were infected with shRNA lentiviral particles produced in HEK293T cells targeted to human MUC1 mRNA, as previously described [18 (link)]. MUC1-specific lentiviral shRNA plasmids were purchased from Sigma-Aldrich (St. Louis, MO).

MCF7 (HTB-22) and MDA-MB468 (HTB-132) breast cancer cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Murine bone marrow derived macrophages (BMDMs) were kindly provided by Dr Luisa Martinez-Pomares, Faculty of Medicine and Health Sciences, University of Nottingham. MCF7 cells were cultured in Eagle’s minimum essential medium (EMEM) (M2279; Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS), 0.01 mg/ml human recombinant insulin, 2 mM l-glutamine, 1 mM sodium pyruvate, 1% minimum essential medium non-essential amino acids. MDA-MB468 cells were cultured in Dulbecco’s modified Eagle medium (DMEM) (D6046; Sigma-Aldrich) supplemented with 10% FBS. BMDMs were cultured in Roswell Park Memorial Institute medium (RPMI) 1640 (R0883; Sigma-Aldrich) supplemented with 10% FBS, 2 mM l-glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin sulphate. All cell lines were incubated in a humidified atmosphere containing 5% CO₂ at 37 °C.

Sunitinib (Selleckchem, Houston, TX) was dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO) and diluted in cell culture media or PBS for in vitro use. Human breast cancer cell lines MDA-MB-231, MDA-MB-468, SKBR-3, MCF-7, and HCC-1419 were obtained from American Type Culture Collection (ATCC, Manassas, VA). The MDA-MB-231, HCC-1419, and MDA-MB-468 cell lines were cultured in RPMI media (Sigma-Aldrich). MCF-7 cell lines were cultured in EMEM media (ATCC). SKBR-3 cells were cultured in McCoy's 5A media (Gibo; Rockville, MD). In all media, 10% fetal bovine serum (Atlantic Biologicals, Miami, FL), 1% Pen/strep (Gibco), 1% sodium pyruvate, 1% NEAA, and 1% glutamine (Gibco) were added and cultured cells were maintained at 37°C and 5% CO₂ humidified atmosphere.

The human breast cancer cell line, MDA-MB-468, was purchased from ATCC (LCG standards, Teddington, UK) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, high glucose, SH30243.01, Cytiva, Marlborough, USA), supplemented with 10% fetal bovine serum (FBS, FBS12A, Capricorn Scientific, Ebsdorfergrund, Germany), and 1% penicillin/streptomycin (15140-122, Gibco, Thermo Fisher Scientific, Waltham, MA, USA), in a humidified atmosphere of 5% CO₂ at 37 °C. The MDA-MB-468 routinely passaged every 2 or 3 days and tested for mycoplasma.
To prepare the bioinks for printing, exponentially growing MDA-MB-468 were trypsinized (T4049, Sigma-Aldrich, St. Louis, MO, USA), and the viable ones were counted on a hemocytometer via trypan blue (15250-061, Gibco) exclusion. The cells were then centrifuged at 1500 rpm for 5 min, and resuspended at 2.5, 7.5, and 20 × 10⁴ cells/μL, in medium with or without 10% glycerol (autoclaved sterile, 40058-ATO, Lach-Ners.r.o.). The resuspended cells were kept on ice until direct printing, normally within one hour.
After printing, the cells (coverslips) were transferred into 24-well plates, and 0.5 mL of complete medium was added to each well.

The BC cell lines were purchased from ATCC (VA, USA) (Additional file 1: Table S1). MCF7, T47D, BT-549 and MDA-MB-231 were cultured in RPMI (Sigma Aldrich, USA), while BT-474, SkBr3, MDA-MB-468, BT-20 and Hs578T were cultured in DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (Sigma Aldrich, USA). The immortalized human mammary epithelial cells hTERT-HME1 (ATCC, USA) were grown in DMEM/F-12 mixture medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (Sigma Aldrich, USA). The human mammary epithelial cells HMEpC were purchased from cell applications (CA, USA) and maintained in defined mammary epithelial cell medium provided by the company (Cell applications, USA).