Dulbecco s modified eagle s high glucose w glutamax 4.5 g l cell culture medium

Self-generated primary equine dermal fibroblasts PriFi1 and PriFi2 and previously isolated primary equine melanoma cells were used for the cell culture experiments. The primary equine melanoma cells MelDuWi belong to the cell culture stock of the Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Germany, while the primary equine melanoma cells eRGO1 were provided by Dr. Barbara Pratscher, Department for Small Animals and Horses, Vetmeduni Vienna, Austria. PriFi1, PriFi2 and MelDuWi were maintained as monolayers in RPMI1640 cell culture medium with stable glutamine (Biochrom GmbH, Berlin, Germany) supplemented with 15% fetal bovine serum (FBS) superior (Biochrom GmbH) and 1% penicillin and streptomycin (10,000 international units (I.U.)/mL / 10,000 μg/mL, Biochrom GmbH) at 37 °C in a humidified atmosphere with 5% CO₂. Melanoma cells eRGO1 were cultured in Dulbecco’s modified Eagle’s high glucose w/Glutamax (4.5 g/L) cell culture medium (GIBCO-Invitrogen, Thermofisher, Darmstadt, Germany) supplemented with 10% FBS superior (Biochrom GmbH) and 1% Antibiotic-Antimycotic (100x; GIBCO-Invitrogen), containing penicillin (10,000 units/mL), streptomycin (10,000 μg/mL) and amphotericin B (25 μg/mL).

All cells used for the experiments originate from different horses. Primary EMM cells (MelDuWi) and primary equine dermal fibroblasts (PriFi1, PriFi2) belong to the cell culture stock of the Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany. The cells were cultured as monolayers at 37°C in a humified atmosphere with 5% CO₂ and maintained in RPMI1640 cell culture medium with stable glutamine (Biochrom GmbH, Berlin, Germany) supplemented with 15% fetal bovine serum superior (Biochrom GmbH) and 1% penicillin and streptomycin (10,000 international units (I.U.)/mL /10,000 μg/mL, Biochrom GmbH). Primary ES cells sRGO1 and sRGO2 (kindly provided by Dr. Sabine Brandt, University of Veterinary Medicine Vienna, Vienna, Austria) and primary EMM cells eRGO1 (kindly provided by Dr. Barbara Pratscher, University of Veterinary Medicine Vienna, Vienna, Austria) were cultured as monolayers at 37°C in a humified atmosphere with 5% CO₂ and kept in Dulbecco’s modified Eagle’s high glucose w/Glutamax (4.5 g/L) cell culture medium (GIBCO-Invitrogen, Thermofisher, Darmstadt, Germany) supplemented with 10% fetal bovine serum superior (Biochrom GmbH) and 1% Antibiotic-Antimycotic (100x; GIBCO-Invitrogen), containing penicillin (10,000 units/mL), streptomycin (10,000 μg/mL) and amphotericin B (25 μg/mL).