Sk n mc

EwS lines (MHH-ES1, SKNMC, and TC-71), were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). A673 was purchased from ATCC (LGC Standards, Teddington, UK). EW7 cells were kindly provided by O. Delattre (Institut Curie, Paris, France). Cells were maintained in a humidified incubator at 37 °C in 5% CO₂ atmosphere in RPMI 1640 (Life Technologies, Darmstadt, Germany) containing 10% heat-inactivated fetal bovine serum (Life Technologies) and antibiotics (Life Technologies). Cell lines were routinely checked for purity (e.g., EWS-FLI1 translocation product, surface antigen, or HLA-phenotype) and mycoplasma contamination as well as for identity by DNA profiling using 8 different and highly polymorphic short tandem repeat (STR) loci (DSMZ, Braunschweig, Germany).

A673 and HEK293T cells were purchased from American Type Culture Collection (ATCC). MHH-ES1, RDES, RH1, SK-ES1, and SK-N-MC cells were provided by the German Collection of Microorganisms and Cell  Cultures (DSMZ). TC-32, TC-71, and CHLA-10 cells were kindly provided by the Children’s Oncology Group (COG) and EW1, EW3, EW7, EW16, EW17, EW18, EW22, EW23, EW24, LAP35, MIC, ORS, POE, STA-ET1, STA-ET8 cells were provided by O. Delattre (Institute Curie, Paris). A673/TR/shEF1 cells were kindly provided by J. Alonso (Madrid, Spain)^{35 (link)}. The SK-N-MC cell line is listed in the database of commonly misidentified cell lines, ICLAC (http://iclac.org/databases/cross-contaminations), as it was initially described to be a neuroblastoma cell line. Indeed, it is a EwS cell line expressing the pathognomonic fusion oncogene EWSR1-FLI1. All cell lines were grown in humidified atmosphere at 37 °C and 5% CO₂. Cells were cultured in RPMI 1640 medium supplemented with stable glutamine (Biochrom), 10% tetracycline-free FCS (Sigma-Aldrich), 100 Uml⁻¹ penicillin (Biochrom), and 100 µg ml⁻¹ streptomycin (Biochrom). Cells were routinely checked by nested PCR for mycoplasma infection, and their purity was confirmed by STR-profiling and, if applicable, by PCR-based detection of specific fusion oncogenes.

SK-N-MC and TC-71 (both ES cell lines) were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ; Braunschweig, Germany). A673 (ES cells) were obtained from ATCC (LGC Standards GmbH, Wesel, Germany). The EW7 ES cell line was obtained from Olivier Delattre, Institut Curie, Paris. The TAP-deficient HLA*A02:01⁺ T2 cell line (somatic cell hybrid) was obtained from P. Cresswell (Yale University School of Medicine, New Haven, CT, USA). The HLA-A*02:01⁻ erythroid leukemia cell line K562 was a gift from A. Knuth and E. Jäger (Krankenhaus Nordwest, Frankfurt, Germany). All cell lines were routinely tested for purity and mycoplasma contamination. Lymphoblastoid cell lines (LCL) were provided by A. Krackhardt (Klinikum rechts der Isar, TU München). HLA genotyping was done at the Labor für Immungenetik und Molekulare Diagnostik (LMU, Munich). Tumor cell lines were cultured in RPMI 1640 supplemented with 10 % fetal calf serum (FCS, Biochrom, Berlin, Germany), 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM L-glutamine (all from Life Technologies). RPMI 1640 medium for LCL and T2 cells was supplemented with 1 mM sodium pyruvate and non-essential amino acids, additionally.

A673 and HEK293 were purchased from the American Type Culture Collection (ATCC). LCL, MHH-ES1, THP-1, SK-N-MC were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ). TC32 was a kind gift from Prof. Poul Sorensen (University of British Columbia, Canada), which was originally purchased from the Childhood Cancer Repository (CCR, Alex’s Lemonade Stand Foundation, Children’s Oncology Group, COG). SB-KMS-KS1 was established at the Children’s Cancer Research Center, Kinderklinik Schwabing (Technical University Munich, Germany). HLA types of utilized cell lines are given in Table 1. IL15-producing NSO cells were a kind gift from Prof. Stanley Riddell (University of Washington School of Medicine), and the packaging cell line (293Vec.) RD114 was kindly provided by Prof. Manuel Caruso (Centre de recherche de Québec, Université Laval). Healthy peripheral blood mononuclear cells (PBMC) were purchased from DRK-Blutspendedienst after informed consent and approval of local government regulatory authorities. Mycoplasma testing was performed regularly (e.g., before in vivo usage, MycoAlert Mycoplasma Detection Kit, Lonza) and cell lines were cultures in accordance with the supplier’s recommendation.

ES lines (SK-N-MC and TC-71), neuroblastoma lines (CHP126, MHH-NB11, SHSY5Y and SIMA) were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany). A673 was purchased from ATCC (LGC Standards, Teddington, UK). Mesenchymal stem cell lines L87 an V54.2 were described previously [33 (link)]. Cells were maintained in a humidified incubator at 37°C in 5-8 % CO₂ atmosphere in RPMI 1640 (Life Technologies, Carlsbad, CA, USA) containing 10 % heat-inactivated fetal bovine serum (Biochrom, Berlin, Germany) and 100 μg/ml gentamicin (Life Technologies). Cell lines were checked routinely for purity (e.g. EWS-FLI1 translocation product, surface antigen or HLA-phenotype) and Mycoplasma contamination.