Gentamicine

Human C33a cervical carcinoma cells (ATCC HTB-31), HCT116 (ATCC CCL-247) and RKO (ATCC CRL-2577) colorectal carcinoma cells, MDA-MB-231 breast carcinoma cells (ATCC HTB-26), A549 lung carcinoma cells (ATCC CCL-185), Panc-1 (ATCC CLR-1469) pancreatic carcinoma and MDCK canine epithelial cells (ATCC CCL-34) were cultured under standard conditions in Dulbecco’s modified Eagle’s medium (BioSera, Nuaille, France) supplemented with 10% fetal calf serum (BioWhittaker, Basel, Switzerland) and gentamicine (Sandoz, Holzkirchen, Germany) in humidified air containing 21% O₂, 5% CO₂ at 37 °C and in hypoxic conditions at anaerobic workstation (1% O₂, 2% H₂, 5% CO₂, 91% N₂, Ruskinn Technology, Bridgend, United Kingdom).

3T3 fibroblasts (American Type Culture Collection) were grown in culture medium containing Dulbecco’s modified Eagle’s medium (DMEM; PAA Laboratories, Pasching, Austria), with 10% fetal bovine serum (FBS; PAA Laboratories) and Primocin TM (100 µg/ml; Lonza, Cologne, Germany). Mesenchymal stem cells (MSCs) were isolated from human umbilical cords obtained from healthy full-term neonates after spontaneous delivery with the informed consent of the donors, using the guidelines approved by the Institutional Ethics Committee at University Hospitals (Pilsen and Prague, Czech Republic). All methods were performed in accordance with the relevant guidelines and regulations of the Institutional Ethics Committee at University Hospitals (Pilsen and Prague, Czech Republic). MSCs were isolated according to previously published protocol^{66 (link)} and expanded in alpha-MEM (EastPort, Prague, Czech Republic), supplemented with 5% platelet lysate (IKEM, Prague, Czech Republic) and gentamicine 10 µg/ml (Sandoz, Prague, Czech Republic). Cells were cultivated at 37 °C in a humidified atmosphere containing 5% CO₂, and the medium was changed twice a week.

HeLa and C33a cervical carcinoma cells, RKO colorectal carcinoma cells and MCF7 breast carcinoma cells were cultured under standard conditions in Dulbecco’s modified Eagle’s medium supplemented with 10% foetal calf serum (Bio Whittaker) and gentamicine (Sandoz) in humidified air containing 21% O₂, 5% CO₂ at 37 °C and in hypoxic conditions at anaerobic workstation (2% O₂, 2% H₂, 5%CO₂, 91% N₂, Ruskinn Technology). All cell lines were purchased from ATCC and regularly tested for Mycoplasma contamination.