Normal human dermal fibroblast cell line

In vitro studies were performed using two models: the Normal Human Dermal Fibroblast (NHDF) cell line (Lonza Group, Basel, Switzerland) and L929 cells, (Sigma-Aldrich, Merck Group, Darmstadt, Germany) (The European Collection of Authenticated Cell Cultures-ECACC). Cells were cultured under standard conditions at 37 °C, 5% CO₂, 95% humidity, in a CO₂ incubator. Cells were always cultured for a minimum of 2 weeks after thawing before starting a series of experiments. Cell cultures were passaged with trypsin/EDTA solution. Cells were counted using a NucleoCounter^® NC-200 automatic cell counter (ChemoMetec A/S, Allerod, Denmark). Cells were cultured in Dulbecco’s modified Eagle medium (DMEM), 10% fetal bovine serum (FBS), penicillin (10,000 U/mL), streptomycin (10 mg/mL), and L-glutamine (200 mM). All culture reagents were purchased from Biological Industries (Biological Industries, Kibbutz Beit-Haemek, Israel).

The Normal Human Dermal Fibroblast (NHDF) cell line from Lonza (Basel, Switzerland) and the L929 cells were purchased from Sigma-Aldrich (Merck Group, Headquarters, Darmstadt, Germany) (The European Collection of Authenticated Cell Cultures—ECACC). All cells were cultured at 37 °C, in 5% CO₂, 95% humidity in a CO₂ incubator. The cells were cultured for a minimum of 2 weeks before the assay. If the confluence exceeded 70% (confluence measurement, JuliBr microscope, NanoEntek, Seoul, Korea), the cell cultures were passaged with trypsin/EDTA solution. The cells were counted with an automatic cell counter NucleoCounter^® NC-200 (ChemoMetec A/S, Allerod, Denmark). If the confluence was less than 70%, the medium was replaced with fresh medium. The cells were cultured in Dulbecco’s modified Eagle medium (DMEM) without phenol red, 10% fetal bovine serum (FBS), penicillin (10,000 U/mL), streptomycin (10 mg/mL), and L-glutamine (200 mM). All culture reagents were purchased from Biological Industries (Beit HaEmek, Israel).