Dulbecco s modified

Human BT-549, A549/KRAS(G12S), and H460/KRAS(Q61H) cells were grown in RPMI1640 medium (ATCC, Manassas, VA, USA). MDA-MB-231 and MDA-MB-468 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Corning, Manassas, VA, USA). BT-20 cells were cultured in Eagle’s Minimum Essential Medium (EMEM) (ATCC). Media were supplanted with 10% heat-inactivated fetal bovine serum (HI-FBS), 100 U/ml penicillin and 100 μg/ml streptomycin. These cells were selected for study based on their comparatively high and low levels of MUC1-C expression (Supplemental Fig. S7). Cells stably expressing MUC1-C were generated as described (60 ). Cells were treated with the MUC1-C inhibitor GO-203 or the control CP-2 peptide (32 (link)). Authentication of cells was performed by short tandem repeat (STR) analysis. Cell were monitored for mycoplasma contamination using the MycoAlert® Mycoplasma Detection Kit (Lonza, Rockland, MA, USA).

Although high viability of cells encapsulated in gels prepared with similar crosslinking protocols have been described in several studies [32 (link),33 (link),35 ], including the use of GelMA, LAP, and light exposure for up to 10 min, we conducted a cell viability assay to assess the impact of the extrusion process on cell viability. NIH/3T3 fibroblasts (ATCC^® CRL-1658™) were cultured in Dulbecco's Modified Eagle's Medium (DMEM) with 4.5 g M⁻¹ glucose, L-glutamine, and sodium pyruvate (Corning, Corning, NY, Cat. No. 10-013) supplemented with 10% heat-inactivated fetal bovine serum and maintained in an incubator at 37 °C and 5% CO2. Cells were subcultured when approximately 80% confluent and detached using trypsin-EDTA (0.25%, phenol red) (ThermoFisher Scientific, Waltham, MA Cat. No. 25200056). Culture medium was changed every 3–4 days. The baseline viability of the cells was assessed using the Trypan Blue Exclusion method prior to encapsulation and the LIVE/DEAD^® assay.

Triple-negative breast cancer cell line MDA-MB-231 was acquired from ATTC. Cells were cultured with Dulbecco’s modified eagle’s medium (DMEM, Corning) and supplemented with 10% v/v HyClone Cosmic Calf Serum (VWR Life Sciences Seradigm), 1% MEM Essential Amino Acids (Quality Biological Inc.), 1% MEM Non-Essential Amino Acids (Quality Biological Inc.), and 50 ng/mL insulin (Sigma- Aldrich, ST. Louis, MO). Cells were grown at 37°C with 5% humidified CO₂ and sub-cultured every 3 days. Abdominal human adipose-derived stem cells (ASCs) were purchased directly from LaCell (New Orleans, LA), and these cells were grown in α-minimum essential medium (α MEM; Gibco, NY), 10% fetal bovine serum (FBS, Atlanta Biologicals), and 1% Antibiotic-Antimycotic (Anti-Anti, Gibco). ASCs were split at 60–70% confluence. The stem cell donor was Caucasian, female, and 24 years old with a body mass index (BMI) of 28. The stem cell medium was changed every third day.