Eagle s minimal essential medium

After obtaining written informed consent, cell lines were established in our laboratory. The MTC-SK cell line (1 (link)) was derived from lymph node metastasis of a 51-year-old central European woman. MTC-SK cells grow as multicellular spheroids in suspension and were maintained in Ham's F12 medium (Biowhittaker, Verviers, Belgium), supplemented with 10% fetal bovine serum (FBS; PAA Laboratories GmbH, Pasching, Austria). HF-SAR fibroblasts (22 (link)) were established from normal skin of a 3-year old male child, cultured in Eagle's minimal essential medium (PAA Laboratories GmbH), supplemented with 10% FBS, in which the cells grew adherently. The cell lines were incubated without antibiotics at 37°C, 5% CO₂ and 95% humidity and were Mycoplasma-free, confirmed with a MycoAlert^®Mycoplasma Detection kit (Cambrex Bioscience, Rockland, ME, USA) and Hoechst 33258 (Sigma-Aldrich, Vienna, Austria). Hoechst stain (50 μl) was added to 50 μl cell suspension at a density of 2×10⁵ (MTC-SK) and 1×10⁵ (HF-SAR) cells/ml and incubated at room temperature for 5 min. The study was approved by the ethics committee of the Medical University of Graz, Austria (18–182 ex 06/07).