Dulbecco s modified eagle s medium high glucose

The colorectal cancer-derived cell lines SW480 and RKO, harboring KRAS and BRAF mutations, respectively, were obtained from the American Type Culture Collection (Manassas, VI, USA). The noncancerous NCM460 cell line derived from normal colon epithelial mucosa was obtained from INCELL’s (San Antonio, TX, USA) [30 (link)]. The cells were grown at 37 °C under a humidified atmosphere containing 5% of CO₂. RKO cell lines were grown in Dulbecco’s Modified Eagle´s Medium High Glucose (Biowest, Nuaillé, France). Both the SW480 and NCM460 cell lines were grown in Roswell Park Memorial Institute 1640 medium with stable glutamine (Biowest, Nuaillé, France). Both mediums were supplemented with 10% fetal bovine serum (v/v) (Biowest, Nuaillé, France) and 1% penicillin/streptomycin (v/v) (Biowest, Nuaillé, France). The cells were subcultured once a week when 80% of confluence was reached and, when necessary, seeded in sterile test plates for the assays. The SW480 and RKO cells were seeded at 1 × 10⁵ cells/mL concentration and the NCM460 at 2 × 10⁵ cells/mL, except for some specific assays.

Bacteria were cultured in three media:1.

Tryptic Soy Broth (Biomaxima, Lublin, Poland), labeled as TSB;

2.

Tryptic Soy Broth (Biomaxima, Lublin, Poland) with 1% of glucose supplementation (w/v; Chempur, Piekary Slaskie, Poland), labeled as TSB + G;

3.

Dulbecco’s Modified Eagle’s Medium High Glucose (Biowest, Riverside, MO, USA; cat no. L0103) supplemented with 10% of fetal bovine serum (Biowest, Riverside, MO, USA), labeled as DMEM.

For experimental purposes, two reference strains from the American Type Culture Collection (ATCC), Staphylococcus aureus 6538 and 33591, and 10 clinical strains isolated from chronic wound infections were chosen. All staphylococcal strains are part of collection of the Department of Pharmaceutical Microbiology and Parasitology of Wroclaw Medical University. Six (five clinical and one reference, 6538) strains were methicillin-susceptible Staphylococcus aureus (MSSA) strains, while another six (five clinical and one reference, 33591) were methicillin-resistant Staphylococcus aureus (MRSA).
All experiments were performed in three different media:

Tryptic soy broth (Biomaxima, Lublin, Poland), later referred to as TSB;

Tryptic soy broth (Biomaxima, Lublin, Poland) supplemented with 1% glucose (w/v; Chempur, Piekary Slaskie, Poland), later referred to as TSB+G;

Dulbecco’s modified Eagle’s medium high glucose (Biowest, Riverside, MO, USA; cat no. L0103) supplemented with 10% of fetal bovine serum (Biowest, Riverside, MO, USA), later referred to as DMEM.