Powerplex esx fast system kit

Human EWS cell lines authentication was executed by short tandem repeat (STR) polymerase chain reaction (PCR) analysis using a PowerPlex ESX Fast System kit (Promega, Madison, WI, USA) and the last control was performed in December 2017. All cell lines were tested for mycoplasma contamination every three months using MycoAlert mycoplasma detection kit (Lonza, Basel, Switzerland). IOR/CAR and LAP-35 cells were immortalized in the Experimental Oncology Laboratory, Rizzoli Orthopaedic Institute of Bologna [26 (link)]. Cultures were grown in Iscove’ Modified Dulbecco’s Medium (IMDM, Thermo Fisher, Milan, Italy) supplemented with 10% of heat-inactivated fetal bovine serum (FBS, Life Technologies, Monza, Italy), 2mM l-glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin (Sigma-Aldrich, Saint Louis, MO, USA) at 37 °C in a humidified atmosphere of 5% CO₂.

EWS cell lines were grown as previously described^{13 (link),40 (link)}. The lines 6647 and TC-71 were kindly provided by T.J. Triche (Children's Hospital, Los Angeles, CA, USA); SK-N-MC, SK-ES-1, and RD-ES were provided by American Type Culture Collection, ATCC (Rockville, MD, USA); IOR/CAR and LAP-35 were previously established in our laboratory; and the A673 sarcoma cell line was provided by Dr. H. Kovar (St. Anna Kinderkrebsforschung, Vienna Austria). Stable CD99-silenced cells were obtained from the TC-71 and IOR/CAR cell lines as previously described^{12 (link),13 (link)}. The cells were cultured in Iscove's modified Dulbecco's medium (IMDM; EuroClone, Milan, Italy) enriched with 10% fetal bovine serum (FBS) (EuroClone) supplemented with 100 U/ml penicillin and 100 mg/ml streptomycin and incubated at 37 °C in a humidified atmosphere at 5% CO₂.
All cell lines were tested for mycoplasma contamination (MycoAlert Mycoplasma Detection Kit, Lonza) and authenticated by short tandem repeat (STR) polymerase chain reaction (PCR) analysis (last control: December 2017) using a PowerPlex ESX Fast System kit (Promega, Madison, WI, USA).