Msc advantage 12 model

Human immortalized keratinocytes—HaCaT (300493; CLS Cell Lines Service GmbH) cells were used in the current study to assess the biological impact of two different cytostatics (Methotrexate and Cetuximab). The culture conditions for HaCaT cell line consists of: Dulbecco’s Modified Eagle’s Medium (DMEM 30-2002™) high glucose (4.5 g/L) media, with 15 mM HEPES (cat. no. H3375), and 2 mM L-glutamine (cat. no. G7513 supplemented with 10% Fetal Calf Serum (cat. no. 12103C) and 1% antibiotic mixture of 100 U/mL penicillin: 100 µg/mL streptomycin (cat. no. P4333), to avoid a possible bacterial infection provided by Sigma-Aldrich, Munich, Germany. The cell cultures were maintained under standard conditions, as described before [19 (link)] by providing an humidified atmosphere enriched in 5% CO2, at 37 °C by means of the Steri-Cycle i160 incubator (Thermo Fisher Scientific, Inc., Waltham, MA, USA). In addition, all the in vitro experiments were performed under sterile conditions by using a biosafety cabinet, MSC Advantage 12 model (Thermo Fisher Scientific, Inc., Waltham, MA, USA).

The cells used in the present study were primary human gingival fibroblasts (HGFs) cells acquired from American Type Culture Collection (ATCC, code no PCS-201-018™). The complete cell growth medium was prepared from Fibroblast Basal Medium (ATCC® PCS-201-030™) supplemented with Fibroblast Growth Kit-Low serum (ATCC® PCS-201-041™) and 0.1% Penicillin-Streptomycin-Amphotericin B Solution (ATCC® PCS-999-002™). All experiments were performed under standard conditions; the cells were grown in a humidified atmosphere with 5% CO₂ and 37 °C in a Steri-Cycle i160 incubator (Thermo Fisher Scientific, Inc., Waltham, MA, USA) and all of the methods were performed under sterile conditions within a biosafety class II cabinet (MSC Advantage 12 model (Thermo Fisher Scientific, Inc., Waltham, MA, USA).